data_7236 #Corrected using PDB structure: 1JTGC # #N.B. (Observed* = Observed shift + Offset correction) # #After reference correction, the following residues still #have a CB chemical shift difference (obs*-pred) greater than 5.0ppm: #NUM AA CS Observed* Predicted # 77 C CB 44.62 37.16 # #After reference correction, the following residues still #have a CO chemical shift difference (obs*-pred) greater than 5.0ppm: #NUM AA CS Observed* Predicted #109 T C 182.02 175.84 # #After reference correction, the following residues still #have a N chemical shift difference (obs*-pred) greater than 10.0ppm: #NUM AA CS Observed* Predicted #234 K N 109.55 119.78 # #After reference correction, the following residues still #have a HN chemical shift difference (obs*-pred) greater than 2.0ppm: #NUM AA CS Observed* Predicted # 69 M H 10.51 8.47 # #The average CS difference between predicted and observed: #HA CA CB CO N HN #N/A -0.27 -0.25 0.05 -0.07 -0.03 # #bmr7236.str.corr chemical shifts have been re-referenced with the following #offsets (these values have been added to the original bmr7236.str file): #HA CA CB CO N HN #N/A -0.26 -0.26 +0.05 -0.07 N/A # #The 95% confidence intervals for the above recommended offsets are: # HA CA CB CO N HN # N/A +/-0.09 +/-0.11 +/-0.10 +/-0.23 +/-0.05 # #The Correlation Coefficients between predicted and observed #chemical shifts are: #HA CA CB CO N HN #N/A 0.981 0.995 0.827 0.853 0.641 # #The RMSD between predicted and observed* (reference #corrected) chemical shifts are: #HA CA CB CO N HN #N/A 0.687 0.734 0.782 1.776 0.356 # ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; NMR Investigation of Tyr105 Mutants in TEM-1 beta-lactamase Suggests Active-Site Dynamical Coupling ; loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Doucet Nicolas . . 2 Savard Pierre-Yves . . 3 Pelletier Joelle N. . 4 Gagne Stephane M. . stop_ _BMRB_accession_number 7236 _BMRB_flat_file_name bmr7236.str _Entry_type new _Submission_date 2006-07-18 _Accession_date 2006-07-18 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "13C chemical shifts" 668 "1H chemical shifts" 247 "15N chemical shifts" 247 stop_ loop_ _Related_BMRB_accession_number _Relationship 6024 "TEM-1 beta-lactamase, E28G mutant" 6357 "TEM-1 beta-lactamase, wild type" 7237 "TEM-1 beta-lactamase, mutant Y105G" 7238 "TEM-1 beta-lactamase, mutant Y105N" 7239 "TEM-1 beta-lactamase, mutant Y105D" stop_ save_ ############################# # Citation for this entry # ############################# save_Citation_1 _Saveframe_category entry_citation _Citation_title ; Backbone dynamics of TEM-1 determined by NMR: evidence for a highly ordered protein. ; _Citation_status published _Citation_type journal _PubMed_ID 16981701 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Savard Pierre-Yves . . 2 Gagne Stephane M. . stop_ _Journal_abbreviation Biochemistry _Journal_volume 45 _Journal_issue 38 _Page_first 11414 _Page_last 11424 _Year 2006 _Details ; The first and second authors contributed equally to this work. ; loop_ _Keyword NMR "TEM-1 beta-lactamase" "15N relaxation" "Enzyme dynamics" stop_ save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name "Mutants Y105W of TEM-1 beta-lactamase" _Abbreviation_common "Mutants Y105W of TEM-1 beta-lactamase" _Enzyme_commission_number 3.5.2.6 loop_ _Mol_system_component_name _Mol_label "TEM-1 Y105W" $TEM-1_beta-lactamase stop_ _System_molecular_weight 29000 _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state "all disulfide bound" loop_ _Biological_function "Cyclic amide hydrolase (beta-lactamase) (E.C. 3.5.2.6)" stop_ _Details ; Four different mutants of the protein TEM-1 in four entries. ; save_ ######################## # Monomeric polymers # ######################## save_TEM-1_beta-lactamase _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common TEM-1 _Name_variant TEM-1 _Abbreviation_common TEM-1 _Mol_thiol_state "all disulfide bound" loop_ _Biological_function beta-lactamase stop_ loop_ _Synonym TEM-1 stop_ ############################## # Polymer residue sequence # ############################## _Residue_count 263 _Mol_residue_sequence ; HPETLVKVKDAEDQLGARVG YIELDLNSGKILESFRPEER FPMMSTFKVLLCGAVLSRVD AGQEQLGRRIHYSQNDLVEW SPVTEKHLTDGMTVRELCSA AITMSDNTAANLLLTTIGGP KELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAA MATTLRKLLTGELLTLASRQ QLIDWMEADKVAGPLLRSAL PAGWFIADKSGAGERGSRGI IAALGPDGKPSRIVVIYTTG SQATMDERNRQIAEIGASLI KHW ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 26 HIS 2 27 PRO 3 28 GLU 4 29 THR 5 30 LEU 6 31 VAL 7 32 LYS 8 33 VAL 9 34 LYS 10 35 ASP 11 36 ALA 12 37 GLU 13 38 ASP 14 39 GLN 15 40 LEU 16 41 GLY 17 42 ALA 18 43 ARG 19 44 VAL 20 45 GLY 21 46 TYR 22 47 ILE 23 48 GLU 24 49 LEU 25 50 ASP 26 51 LEU 27 52 ASN 28 53 SER 29 54 GLY 30 55 LYS 31 56 ILE 32 57 LEU 33 58 GLU 34 59 SER 35 60 PHE 36 61 ARG 37 62 PRO 38 63 GLU 39 64 GLU 40 65 ARG 41 66 PHE 42 67 PRO 43 68 MET 44 69 MET 45 70 SER 46 71 THR 47 72 PHE 48 73 LYS 49 74 VAL 50 75 LEU 51 76 LEU 52 77 CYS 53 78 GLY 54 79 ALA 55 80 VAL 56 81 LEU 57 82 SER 58 83 ARG 59 84 VAL 60 85 ASP 61 86 ALA 62 87 GLY 63 88 GLN 64 89 GLU 65 90 GLN 66 91 LEU 67 92 GLY 68 93 ARG 69 94 ARG 70 95 ILE 71 96 HIS 72 97 TYR 73 98 SER 74 99 GLN 75 100 ASN 76 101 ASP 77 102 LEU 78 103 VAL 79 104 GLU 80 105 TRP 81 106 SER 82 107 PRO 83 108 VAL 84 109 THR 85 110 GLU 86 111 LYS 87 112 HIS 88 113 LEU 89 114 THR 90 115 ASP 91 116 GLY 92 117 MET 93 118 THR 94 119 VAL 95 120 ARG 96 121 GLU 97 122 LEU 98 123 CYS 99 124 SER 100 125 ALA 101 126 ALA 102 127 ILE 103 128 THR 104 129 MET 105 130 SER 106 131 ASP 107 132 ASN 108 133 THR 109 134 ALA 110 135 ALA 111 136 ASN 112 137 LEU 113 138 LEU 114 139 LEU 115 140 THR 116 141 THR 117 142 ILE 118 143 GLY 119 144 GLY 120 145 PRO 121 146 LYS 122 147 GLU 123 148 LEU 124 149 THR 125 150 ALA 126 151 PHE 127 152 LEU 128 153 HIS 129 154 ASN 130 155 MET 131 156 GLY 132 157 ASP 133 158 HIS 134 159 VAL 135 160 THR 136 161 ARG 137 162 LEU 138 163 ASP 139 164 ARG 140 165 TRP 141 166 GLU 142 167 PRO 143 168 GLU 144 169 LEU 145 170 ASN 146 171 GLU 147 172 ALA 148 173 ILE 149 174 PRO 150 175 ASN 151 176 ASP 152 177 GLU 153 178 ARG 154 179 ASP 155 180 THR 156 181 THR 157 182 MET 158 183 PRO 159 184 ALA 160 185 ALA 161 186 MET 162 187 ALA 163 188 THR 164 189 THR 165 190 LEU 166 191 ARG 167 192 LYS 168 193 LEU 169 194 LEU 170 195 THR 171 196 GLY 172 197 GLU 173 198 LEU 174 199 LEU 175 200 THR 176 201 LEU 177 202 ALA 178 203 SER 179 204 ARG 180 205 GLN 181 206 GLN 182 207 LEU 183 208 ILE 184 209 ASP 185 210 TRP 186 211 MET 187 212 GLU 188 213 ALA 189 214 ASP 190 215 LYS 191 216 VAL 192 217 ALA 193 218 GLY 194 219 PRO 195 220 LEU 196 221 LEU 197 222 ARG 198 223 SER 199 224 ALA 200 225 LEU 201 226 PRO 202 227 ALA 203 228 GLY 204 229 TRP 205 230 PHE 206 231 ILE 207 232 ALA 208 233 ASP 209 234 LYS 210 235 SER 211 236 GLY 212 237 ALA 213 238 GLY 214 239 GLU 215 240 ARG 216 241 GLY 217 242 SER 218 243 ARG 219 244 GLY 220 245 ILE 221 246 ILE 222 247 ALA 223 248 ALA 224 249 LEU 225 250 GLY 226 251 PRO 227 252 ASP 228 253 GLY 229 254 LYS 230 255 PRO 231 256 SER 232 257 ARG 233 258 ILE 234 259 VAL 235 260 VAL 236 261 ILE 237 262 TYR 238 263 THR 239 264 THR 240 265 GLY 241 266 SER 242 267 GLN 243 268 ALA 244 269 THR 245 270 MET 246 271 ASP 247 272 GLU 248 273 ARG 249 274 ASN 250 275 ARG 251 276 GLN 252 277 ILE 253 278 ALA 254 279 GLU 255 280 ILE 256 281 GLY 257 282 ALA 258 283 SER 259 284 LEU 260 285 ILE 261 286 LYS 262 287 HIS 263 288 TRP stop_ _Sequence_homology_query_date 2007-03-02 _Sequence_homology_query_revised_last_date 2007-01-18 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value BMRB 6024 "TEM-1 beta-lactamase" 100.00 263 99 100 10e-147 BMRB 6357 "TEM-1 beta-lactamase" 100.00 263 100 100 10e-147 BMRB 7237 TEM-1 100.00 263 100 100 10e-147 BMRB 7238 TEM-1 100.00 263 100 100 10e-147 BMRB 7239 TEM-1 100.00 263 100 100 10e-147 PDB 1ERM "A Chain A, X-Ray Crystal Structure Of Tem-1Beta Lactamase In Complex With A Designed Boronic AcidInhibitor (1r)-1-Acetamido-2- (3-Carboxyphenyl)ethaneBoronic Acid" 100.00 263 99 100 10e-147 PDB 1ERO "A Chain A, X-Ray Crystal Structure Of Tem-1Beta Lactamase In Complex With A Designed Boronic AcidInhibitor (1r)-2-Phenylacetamido-2-(3-Carboxyphenyl)ethyl Boronic Acid" 100.00 263 100 100 10e-147 PDB 1ERQ "A Chain A, X-Ray Crystal Structure Of Tem-1Beta Lactamase In Complex With A Designed Boronic AcidInhibitor (1r)-1-Acetamido-2-(3-Carboxy-2-Hydroxyphenyl)ethyl Boronic Acid" 100.00 263 100 100 10e-147 PDB 1ESU "A Chain A, S235a Mutant Of Tem1 Beta-Lactamase" 100.00 263 99 100 10e-147 PDB 1FQG "A Chain A, Molecular Structure Of TheAcyl-Enzyme Intermediate In Tem- 1 Beta-Lactamase" 100.00 263 99 100 10e-147 PDB 1JTD "A Chain A, Crystal Structure Of Beta-LactamaseInhibitor Protein-Ii In Complex With Tem-1Beta-Lactamase" 100.00 263 99 100 10e-147 PDB 1JVJ "A Chain A, Crystal Structure Of N132a MutantOf Tem-1 Beta-Lactamase In Complex With AN-Formimidoyl-Thienamycine" 100.00 263 99 100 10e-147 PDB 1JWP "A Chain A, Structure Of M182t Mutant Of Tem-1Beta-Lactamase" 100.00 263 99 100 10e-147 PDB 1JWV "A Chain A, Crystal Structure Of G238a MutantOf Tem-1 Beta-Lactamase In Complex With A Boronic AcidInhibitor (Sefb4)" 100.00 263 99 100 10e-147 PDB 1LHY "A Chain A, Crystal Structure Of Tem-30Beta-Lactamase At 2.0 Angstrom" 100.00 263 99 100 10e-147 PDB 1LI9 "A Chain A, Crystal Structure Of Tem-34Beta-Lactamase At 1.5 Angstrom" 100.00 263 99 100 10e-147 PDB 1M40 "A Chain A, Ultra High Resolution CrystalStructure Of Tem-1" 100.00 263 99 100 10e-147 PDB 1NXY "A Chain A, Crystal Structure Of The ComplexBetween M182t Mutant Of Tem-1 And A Boronic AcidInhibitor (Sm2)" 100.00 263 99 100 10e-147 PDB 1NY0 "A Chain A, Crystal Structure Of The ComplexBetween M182t Mutant Of Tem-1 And A Boronic AcidInhibitor (Nbf)" 100.00 263 99 100 10e-147 PDB 1NYM "A Chain A, Crystal Structure Of The ComplexBetween M182t Mutant Of Tem-1 And A Boronic AcidInhibitor (Cxb)" 100.00 263 99 100 10e-147 PDB 1NYY "A Chain A, Crystal Structure Of The ComplexBetween M182t Mutant Of Tem-1 And A Boronic AcidInhibitor (105)" 100.00 263 99 100 10e-147 PDB 1PZO "A Chain A, Tem-1 Beta-Lactamase In ComplexWith A Novel, Core- Disrupting, Allosteric Inhibitor" 100.00 263 99 100 10e-147 PDB 1PZP "A Chain A, Tem-1 Beta-Lactamase In ComplexWith A Novel, Core- Disrupting, Allosteric Inhibitor" 100.00 263 99 100 10e-147 PDB 1S0W "A Chain A, 1b Lactamse B LACTAMASE INHIBITOR" 100.00 263 100 100 10e-147 PDB 1XPB "Chain , Structure Of Beta-Lactamase Tem1" 100.00 263 100 100 10e-147 PDB 1YT4 "A Chain A, Crystal Structure Of Tem-76Beta-Lactamase At 1.4 Angstrom Resolution" 100.00 263 99 100 10e-147 PDB 1AXB "Chain , Tem-1 Beta-Lactamase FromEscherichia Coli Inhibited With An Acylation TransitionState Analog" 100.00 263 99 100 10e-147 PDB 1BT5 "A Chain A, Crystal Structure Of The ImipenemInhibited Tem-1 Beta- Lactamase From Escherichia Coli" 100.00 263 99 100 10e-147 PDB 1BTL "Chain , Beta-Lactamase Tem1 (E.C.3.5.2.6)" 100.00 263 99 100 10e-147 PDB 1CK3 "A Chain A, N276d Mutant Of Escherichia ColiTem-1 Beta-Lactamase" 100.00 263 98 100 1e-145 PDB 1HTZ "A Chain A, Crystal Structure Of Tem52Beta-Lactamase" 100.00 263 98 99 1e-145 PDB 1JTG "A Chain A, Crystal Structure Of Tem-1Beta-Lactamase BETA-Lactamase Inhibitor Protein Complex" 100.00 263 99 100 10e-147 PDB 1JWZ "A Chain A, Crystal Structure Of Tem-64Beta-Lactamase In Complex With A Boronic Acid Inhibitor(105)" 100.00 263 98 99 1e-145 PDB 1LI0 "A Chain A, Crystal Structure Of Tem-32Beta-Lactamase At 1.6 Angstrom" 100.00 263 99 100 1e-145 PDB 1TEM "Chain , 6 Alpha Hydroxymethyl PenicilloicAcid Acylated On The Tem-1 Beta-Lactamase FromEscherichia Coli" 100.00 263 99 100 10e-147 PDB 1XXM "A Chain A, The Modular Architecture OfProtein-Protein Binding Site" 100.00 263 99 99 1e-145 PDB 2B5R "A Chain A, 1b Lactamase B LACTAMASE INHIBITOR" 100.00 263 99 99 1e-145 PDB 1ZG4 "A Chain A, Tem1 Beta Lactamase" 91.96 286 99 100 10e-147 PDB 1ZG6 "A Chain A, Tem1 Beta Lactamase Mutant S70g" 91.96 286 98 99 1e-145 DBJ BAA00795.1 "ampicillin resistance protein [syntheticconstruct]" 91.96 286 100 100 10e-147 DBJ BAA14388.1 "Ap resistance protein [syntheticconstruct]" 91.96 286 100 100 10e-147 DBJ BAA21464.1 "beta-lactamase [Cloning vector pMW118]" 91.96 286 100 100 10e-147 DBJ BAA21466.1 "beta-lactamase [Cloning vector pMW119]" 91.96 286 100 100 10e-147 DBJ BAA93577.1 "beta-lactamase [Cloning vector pGFPTA]" 91.96 286 100 100 10e-147 EMBL CAA23886.1 "beta-lactamase [Escherichia coli]" 91.96 286 100 100 10e-147 EMBL CAA47409.1 "bla gene [synthetic construct]" 91.96 286 100 100 10e-147 EMBL CAA47411.1 "bla gene from [synthetic construct]" 91.96 286 100 100 10e-147 EMBL CAA86303.1 "Beta-Lactamase [Pichia pastoris]" 91.96 286 100 100 10e-147 EMBL CAA48214.1 "beta-lactamase,human IgG3 hinge fusion[synthetic construct]" 73.88 356 100 100 10e-147 GenBank AAD34466.1 "AF136442_2 beta-lactamase [Cloning vectorpScosBC1]" 91.96 286 100 100 10e-147 GenBank AAF01047.1 "AF188200_1 beta-lactamase variant TEM-1D[Escherichia coli]" 91.96 286 100 100 10e-147 GenBank AAF01198.1 "AF179892_2 beta-lactamase [Cloning vectorpLM3]" 91.96 286 100 100 10e-147 GenBank ABD61074.1 "TEM-1 beta lactamase gene [Neisseriagonorrhoeae]" 91.96 286 100 100 10e-147 GenBank AAQ93652.1 "RDfusion [Ribosome display vector pRDV]" 45.27 581 100 100 10e-147 PIR S60310 "extended spectrum beta-lactamase CAZ-2 -Klebsiella pneumoniae" 91.96 286 98 99 1e-144 PIR S60312 "extended spectrum beta-lactamase CAZ-7 -Klebsiella pneumoniae" 91.96 286 98 100 1e-145 PIR T51301 "beta-lactamase (EC 3.5.2.6) - fission yeast(Schizosaccharomyces pombe)" 91.96 286 99 100 10e-147 PRF 2018199A "beta lactamase IRT-4" 91.96 286 100 100 10e-147 REF NP_052173.1 "beta-lactamase [Neisseria gonorrhoeae]" 91.96 286 100 100 10e-147 REF NP_569411.1 "beta-lactamase [Salmonella entericasubsp. enterica serovar Typhi str. CT18]" 91.96 286 100 100 10e-147 REF NP_608310.1 "TEM beta-lactamase [Klebsiellapneumoniae]" 91.96 286 100 100 10e-147 REF NP_775035.1 "BlaTEM1 [Citrobacter freundii]" 91.96 286 100 100 10e-147 REF NP_943295.1 "beta-lactamase [Escherichia coli]" 91.96 286 100 100 10e-147 SWISS-PROT P62593 "BLAT_ECOLI Beta-lactamase TEM precursor(TEM-1) (TEM-2) (TEM-3) (TEM-4) (TEM-5) (TEM-6)(TEM-8/CAZ-2) (TEM-16/CAZ-7) (TEM-24/CAZ-6) (IRT-4)(Penicillinase)" 91.96 286 100 100 10e-147 SWISS-PROT P62594 "BLAT_SALTI Beta-lactamase TEM precursor(Penicillinase)" 91.96 286 100 100 10e-147 stop_ save_ ######################################## # Molecular bond linkage definitions # ######################################## save_crosslink_bonds _Saveframe_category crosslink_bonds loop_ _Bond_order _Bond_type _Atom_one_mol_system_component_name _Atom_one_residue_seq_code _Atom_one_residue_label _Atom_one_atom_name _Atom_two_mol_system_component_name _Atom_two_residue_seq_code _Atom_two_residue_label _Atom_two_atom_name single disulfide "TEM-1 Y105W" 52 CYS SG "TEM-1 Y105W" 98 CYS SG stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $TEM-1_beta-lactamase "E. coli" 562 Eubacteria ? Escherichia coli stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Variant _Vector_name $TEM-1_beta-lactamase "recombinant technology" "E. coli" Escherichia coli BL21 DE3 ? stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details "0.8 mM, pH 6.6 for all Y105X mutants" loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $TEM-1_beta-lactamase 0.8 mM "[U-13C; U-15N]" imidazole 4 mM ? DSS 0.1 mM ? H2O 90 % ? D2O 10 % ? stop_ save_ ############################ # Computer software used # ############################ save_software_1 _Saveframe_category software _Name Vnmr _Version 6.1c loop_ _Vendor _Address _Electronic_address Varian ; Varian Inc. 3120 Hansen Way Palo Alto, CA 94304-1030 USA ; http://www.varianinc.com stop_ loop_ _Task "spectra acquisition" stop_ save_ save_software_2 _Saveframe_category software _Name nmrPipe _Version 2.3 loop_ _Vendor _Address _Electronic_address "Frank Delaglio (Ad Bax)" http://spin.niddk.nih.gov/NMRPipe/ http://spin.niddk.nih.gov/bax/people/delaglio.html stop_ loop_ _Task "Transform/process data" stop_ save_ save_software_3 _Saveframe_category software _Name nmrView _Version 5.2.2 loop_ _Vendor _Address _Electronic_address "One moon scientific" http://www.onemoonscientific.com/index.html http://www.onemoonscientific.com/index.html stop_ loop_ _Task "analyze data" stop_ save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_600MHz_spectrometer _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model INOVA _Field_strength 600 _Details "with cryoprobe" save_ ############################# # NMR applied experiments # ############################# save_NMR_experiment_list _Saveframe_category NMR_applied_experiment _Experiment_name ; 15N-HSQC 3D-HNCO 3D-HN(CO)CA 3D-CBCA(CO)NH ; save_ save_15N-HSQC _Saveframe_category NMR_applied_experiment _Experiment_name 15N-HSQC _Sample_label $sample_1 save_ save_3D-HNCO _Saveframe_category NMR_applied_experiment _Experiment_name 3D-HNCO _Sample_label $sample_1 save_ save_3D-HN(CO)CA _Saveframe_category NMR_applied_experiment _Experiment_name 3D-HN(CO)CA _Sample_label $sample_1 save_ save_3D-CBCA(CO)NH _Saveframe_category NMR_applied_experiment _Experiment_name 3D-CBCA(CO)NH _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_conditions_1 _Saveframe_category sample_conditions _Details ; The enzymes were dissolved to a concentration of 0.8 mM in a 90 % H2O:10 % D2O solution containing 4 mM imidazole and 0.1 mM DSS (pH = 6.6). All NMR experiments were performed at 30?C on a Varian INOVA 600 spectrometer operating at a proton frequency of 599.739 MHz equipped with a z-axis gradient and a triple resonance cryoprobe. ; loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 6.6 0.2 pH temperature 303 0.2 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_referencing _Saveframe_category chemical_shift_reference loop_ _Mol_common_name _Atom_type _Atom_isotope_number _Atom_group _Chem_shift_units _Chem_shift_value _Reference_method _Reference_type _External_reference_sample_geometry _External_reference_location _External_reference_axis _Indirect_shift_ratio DSS H 1 "methyl protons" ppm 0 direct internal ? ? ? 1.0 DSS N 15 "methyl protons" ppm 0 indirect internal ? ? ? 0.101329118 DSS C 13 "methyl protons" ppm 0 indirect internal ? ? ? 0.25144953 stop_ save_ ################################### # Assigned chemical shift lists # ################################### ################################################################### # Chemical Shift Ambiguity Index Value Definitions # # # # Index Value Definition # # # # 1 Unique (geminal atoms and geminal methyl # # groups with identical chemical shifts # # are assumed to be assigned to # # stereospecific atoms) # # 2 Ambiguity of geminal atoms or geminal methyl # # proton groups # # 3 Aromatic atoms on opposite sides of # # symmetrical rings (e.g. Tyr HE1 and HE2 # # protons) # # 4 Intraresidue ambiguities (e.g. Lys HG and # # HD protons or Trp HZ2 and HZ3 protons) # # 5 Interresidue ambiguities (Lys 12 vs. Lys 27) # # 9 Ambiguous, specific ambiguity not defined # # # ################################################################### save_chem_shift_list_1 _Saveframe_category assigned_chemical_shifts _Details "same for all Y105X mutants" loop_ _Experiment_label $15N-HSQC $3D-HNCO $3D-HN(CO)CA $3D-CBCA(CO)NH stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $conditions_1 _Chem_shift_reference_set_label $chemical_shift_referencing _Mol_system_component_name "TEM-1 Y105W" loop_ _Atom_shift_assign_ID _Residue_seq_code _Residue_label _Atom_name _Atom_type _Chem_shift_value _Chem_shift_value_error _Chem_shift_ambiguity_code 1 27 PRO C C 179.43 0.2 1 2 27 PRO CA C 65.35 0.2 1 3 27 PRO CB C 32.14 0.2 1 4 28 GLU H H 11.10 0.02 1 5 28 GLU C C 179.64 0.2 1 6 28 GLU CA C 59.39 0.2 1 8 28 GLU N N 121.71 0.15 1 9 29 THR H H 7.81 0.02 1 10 29 THR C C 176.13 0.2 1 11 29 THR CA C 64.91 0.2 1 12 29 THR CB C 67.41 0.2 1 13 29 THR N N 118.87 0.15 1 14 30 LEU H H 7.07 0.02 1 15 30 LEU C C 179.06 0.2 1 16 30 LEU CA C 57.23 0.2 1 17 30 LEU CB C 40.55 0.2 1 18 30 LEU N N 120.16 0.15 1 19 31 VAL H H 7.54 0.02 1 20 31 VAL C C 178.50 0.2 1 21 31 VAL CA C 66.61 0.2 1 22 31 VAL CB C 31.17 0.2 1 23 31 VAL N N 119.69 0.15 1 24 32 LYS H H 7.32 0.02 1 25 32 LYS C C 178.44 0.2 1 26 32 LYS CA C 56.88 0.2 1 27 32 LYS CB C 30.15 0.2 1 28 32 LYS N N 119.43 0.15 1 29 33 VAL H H 8.34 0.02 1 30 33 VAL C C 176.86 0.2 1 31 33 VAL CA C 67.01 0.2 1 32 33 VAL CB C 30.75 0.2 1 33 33 VAL N N 122.96 0.15 1 34 34 LYS H H 7.73 0.02 1 35 34 LYS C C 179.41 0.2 1 36 34 LYS CA C 59.68 0.2 1 37 34 LYS CB C 31.68 0.2 1 38 34 LYS N N 118.52 0.15 1 39 35 ASP H H 8.35 0.02 1 43 35 ASP N N 120.50 0.15 1 44 36 ALA H H 8.55 0.02 1 45 36 ALA C C 179.65 0.2 1 46 36 ALA CA C 55.23 0.2 1 47 36 ALA CB C 18.13 0.2 1 49 37 GLU H H 7.97 0.02 1 50 37 GLU C C 180.53 0.2 1 51 37 GLU CA C 60.83 0.2 1 52 37 GLU CB C 28.92 0.2 1 53 37 GLU N N 117.62 0.15 1 54 38 ASP H H 7.94 0.02 1 55 38 ASP C C 179.20 0.2 1 56 38 ASP CA C 56.99 0.2 1 57 38 ASP CB C 40.75 0.2 1 58 38 ASP N N 119.90 0.15 1 59 39 GLN H H 8.89 0.02 1 60 39 GLN C C 179.23 0.2 1 61 39 GLN CA C 58.29 0.2 1 62 39 GLN CB C 28.34 0.2 1 63 39 GLN N N 118.42 0.15 1 64 40 LEU H H 8.76 0.02 1 68 40 LEU N N 114.32 0.15 1 69 41 GLY H H 8.12 0.02 1 70 41 GLY C C 174.06 0.2 1 71 41 GLY CA C 47.12 0.2 1 73 42 ALA H H 7.76 0.02 1 74 42 ALA C C 174.95 0.2 1 75 42 ALA CA C 50.52 0.2 1 77 42 ALA N N 119.55 0.15 1 78 43 ARG H H 7.65 0.02 1 82 43 ARG N N 115.40 0.15 1 83 44 VAL H H 10.70 0.02 1 84 44 VAL C C 172.74 0.2 1 88 45 GLY H H 8.91 0.02 1 89 45 GLY C C 172.54 0.2 1 90 45 GLY CA C 43.76 0.2 1 91 45 GLY N N 111.30 0.15 1 92 46 TYR H H 9.44 0.02 1 93 46 TYR C C 174.10 0.2 1 94 46 TYR CA C 56.78 0.2 1 95 46 TYR CB C 44.99 0.2 1 96 46 TYR N N 124.01 0.15 1 97 47 ILE H H 8.06 0.02 1 98 47 ILE C C 171.54 0.2 1 99 47 ILE CA C 59.11 0.2 1 101 47 ILE N N 125.42 0.15 1 102 48 GLU H H 7.84 0.02 1 103 48 GLU C C 175.08 0.2 1 104 48 GLU CA C 54.14 0.2 1 106 48 GLU N N 123.50 0.15 1 107 49 LEU H H 9.72 0.02 1 108 49 LEU C C 175.70 0.2 1 109 49 LEU CA C 52.96 0.2 1 110 49 LEU CB C 46.99 0.2 1 111 49 LEU N N 126.80 0.15 1 112 50 ASP H H 9.07 0.02 1 113 50 ASP C C 176.85 0.2 1 114 50 ASP CA C 55.32 0.2 1 115 50 ASP CB C 42.87 0.2 1 116 50 ASP N N 126.53 0.15 1 117 51 LEU H H 8.26 0.02 1 118 51 LEU C C 177.63 0.2 1 119 51 LEU CA C 58.52 0.2 1 120 51 LEU CB C 42.87 0.2 1 121 51 LEU N N 128.85 0.15 1 122 52 ASN H H 8.71 0.02 1 123 52 ASN C C 177.03 0.2 1 124 52 ASN CA C 56.14 0.2 1 126 52 ASN N N 116.55 0.15 1 127 53 SER H H 8.58 0.02 1 128 53 SER C C 176.56 0.2 1 129 53 SER CA C 58.79 0.2 1 130 53 SER CB C 65.52 0.2 1 131 53 SER N N 112.81 0.15 1 132 54 GLY H H 8.02 0.02 1 133 54 GLY C C 172.42 0.2 1 134 54 GLY CA C 45.38 0.2 1 135 54 GLY N N 112.19 0.15 1 136 55 LYS H H 7.71 0.02 1 137 55 LYS C C 176.18 0.2 1 138 55 LYS CA C 56.33 0.2 1 139 55 LYS CB C 33.53 0.2 1 140 55 LYS N N 119.35 0.15 1 141 56 ILE H H 8.69 0.02 1 142 56 ILE C C 176.19 0.2 1 143 56 ILE CA C 62.43 0.2 1 145 56 ILE N N 124.84 0.15 1 146 57 LEU H H 8.96 0.02 1 147 57 LEU C C 177.13 0.2 1 148 57 LEU CA C 55.62 0.2 1 149 57 LEU CB C 41.91 0.2 1 150 57 LEU N N 130.41 0.15 1 151 58 GLU H H 7.57 0.02 1 152 58 GLU C C 174.70 0.2 1 153 58 GLU CA C 54.79 0.2 1 154 58 GLU CB C 34.08 0.2 1 155 58 GLU N N 115.90 0.15 1 156 59 SER H H 9.03 0.02 1 157 59 SER C C 172.08 0.2 1 158 59 SER CA C 57.72 0.2 1 159 59 SER CB C 66.90 0.2 1 160 59 SER N N 115.50 0.15 1 161 60 PHE H H 9.57 0.02 1 162 60 PHE C C 173.35 0.2 1 163 60 PHE CA C 59.66 0.2 1 165 60 PHE N N 122.33 0.15 1 166 61 ARG H H 8.78 0.02 1 167 61 ARG N N 121.73 0.15 1 168 62 PRO C C 177.58 0.2 1 169 62 PRO CA C 65.03 0.2 1 170 62 PRO CB C 32.43 0.2 1 171 63 GLU H H 8.51 0.02 1 172 63 GLU C C 175.13 0.2 1 173 63 GLU CA C 54.75 0.2 1 174 63 GLU CB C 28.88 0.2 1 175 63 GLU N N 116.46 0.15 1 176 64 GLU H H 6.87 0.02 1 177 64 GLU C C 174.94 0.2 1 178 64 GLU CA C 54.77 0.2 1 179 64 GLU CB C 32.09 0.2 1 180 64 GLU N N 117.25 0.15 1 181 65 ARG H H 8.33 0.02 1 182 65 ARG C C 177.30 0.2 1 183 65 ARG CA C 55.70 0.2 1 184 65 ARG CB C 31.71 0.2 1 185 65 ARG N N 117.15 0.15 1 186 66 PHE H H 8.75 0.02 1 187 66 PHE N N 117.81 0.15 1 188 67 PRO C C 176.20 0.2 1 189 67 PRO CA C 62.50 0.2 1 190 68 MET H H 7.81 0.02 1 191 68 MET C C 179.72 0.2 1 192 68 MET CA C 58.17 0.2 1 193 68 MET N N 119.31 0.15 1 194 69 MET H H 10.54 0.02 1 195 69 MET N N 120.39 0.15 1 196 70 SER C C 176.14 0.2 1 197 71 THR H H 7.92 0.02 1 198 71 THR C C 175.29 0.2 1 199 71 THR CA C 65.70 0.2 1 200 71 THR N N 112.06 0.15 1 201 72 PHE H H 7.75 0.02 1 202 72 PHE C C 176.66 0.2 1 203 72 PHE CA C 59.21 0.2 1 204 72 PHE CB C 39.32 0.2 1 205 72 PHE N N 116.77 0.15 1 206 73 LYS H H 7.44 0.02 1 207 73 LYS C C 177.43 0.2 1 208 73 LYS CA C 59.78 0.2 1 209 73 LYS N N 122.25 0.15 1 210 74 VAL H H 6.64 0.02 1 211 74 VAL C C 176.05 0.2 1 212 74 VAL CA C 66.31 0.2 1 213 74 VAL N N 115.50 0.15 1 214 75 LEU H H 6.68 0.02 1 215 75 LEU C C 178.11 0.2 1 216 75 LEU CA C 57.19 0.2 1 217 75 LEU CB C 40.37 0.2 1 218 75 LEU N N 117.74 0.15 1 219 76 LEU H H 8.04 0.02 1 220 76 LEU C C 177.28 0.2 1 221 76 LEU CA C 57.26 0.2 1 222 76 LEU CB C 42.27 0.2 1 223 76 LEU N N 116.73 0.15 1 224 77 CYS H H 7.50 0.02 1 225 77 CYS C C 175.71 0.2 1 226 77 CYS CA C 63.60 0.2 1 227 77 CYS CB C 44.62 0.2 1 228 77 CYS N N 113.74 0.15 1 229 78 GLY H H 8.40 0.02 1 230 78 GLY C C 173.18 0.2 1 231 78 GLY CA C 47.63 0.2 1 232 78 GLY N N 110.96 0.15 1 233 79 ALA H H 7.85 0.02 1 234 79 ALA C C 181.05 0.2 1 235 79 ALA CA C 54.82 0.2 1 236 79 ALA CB C 16.40 0.2 1 237 79 ALA N N 124.60 0.15 1 238 80 VAL H H 8.03 0.02 1 241 80 VAL N N 119.36 0.15 1 242 81 LEU H H 8.51 0.02 1 243 81 LEU C C 178.29 0.2 1 247 82 SER H H 8.20 0.02 1 248 82 SER C C 178.05 0.2 1 249 82 SER CA C 61.66 0.2 1 250 82 SER N N 115.67 0.15 1 251 83 ARG H H 7.52 0.02 1 252 83 ARG C C 179.46 0.2 1 253 83 ARG CA C 59.17 0.2 1 254 83 ARG CB C 30.25 0.2 1 255 83 ARG N N 121.88 0.15 1 256 84 VAL H H 8.32 0.02 1 257 84 VAL C C 180.90 0.2 1 258 84 VAL CA C 64.95 0.2 1 259 84 VAL CB C 30.82 0.2 1 260 84 VAL N N 124.38 0.15 1 261 85 ASP H H 9.11 0.02 1 262 85 ASP C C 177.83 0.2 1 263 85 ASP CA C 57.10 0.2 1 264 85 ASP CB C 40.73 0.2 1 265 85 ASP N N 123.38 0.15 1 266 86 ALA H H 7.54 0.02 1 267 86 ALA C C 177.98 0.2 1 268 86 ALA CA C 52.03 0.2 1 269 86 ALA N N 118.52 0.15 1 270 87 GLY H H 8.23 0.02 1 271 87 GLY C C 174.98 0.2 1 272 87 GLY CA C 45.64 0.2 1 273 87 GLY N N 107.64 0.15 1 274 88 GLN H H 8.42 0.02 1 275 88 GLN C C 173.57 0.2 1 276 88 GLN CA C 55.40 0.2 1 277 88 GLN CB C 29.85 0.2 1 278 88 GLN N N 117.25 0.15 1 279 89 GLU H H 7.38 0.02 1 280 89 GLU C C 172.75 0.2 1 281 89 GLU CA C 52.63 0.2 1 282 89 GLU CB C 33.60 0.2 1 283 89 GLU N N 117.98 0.15 1 284 90 GLN H H 10.09 0.02 1 285 90 GLN C C 176.10 0.2 1 286 90 GLN CA C 54.69 0.2 1 288 90 GLN N N 125.16 0.15 1 289 91 LEU H H 9.08 0.02 1 290 91 LEU C C 177.39 0.2 1 291 91 LEU CA C 57.44 0.2 1 292 91 LEU CB C 41.36 0.2 1 293 91 LEU N N 123.05 0.15 1 294 92 GLY H H 8.39 0.02 1 295 92 GLY C C 173.93 0.2 1 296 92 GLY CA C 44.26 0.2 1 297 92 GLY N N 102.03 0.15 1 298 93 ARG H H 7.36 0.02 1 299 93 ARG C C 174.77 0.2 1 300 93 ARG CA C 57.77 0.2 1 302 93 ARG N N 122.48 0.15 1 303 94 ARG H H 8.54 0.02 1 304 94 ARG C C 174.61 0.2 1 305 94 ARG CA C 56.30 0.2 1 306 94 ARG CB C 31.35 0.2 1 307 94 ARG N N 127.26 0.15 1 308 95 ILE H H 9.14 0.02 1 309 95 ILE C C 174.53 0.2 1 310 95 ILE CA C 60.21 0.2 1 311 95 ILE CB C 38.97 0.2 1 312 95 ILE N N 130.25 0.15 1 313 96 HIS H H 8.47 0.02 1 314 96 HIS C C 174.24 0.2 1 315 96 HIS CA C 55.25 0.2 1 316 96 HIS CB C 30.85 0.2 1 317 96 HIS N N 124.56 0.15 1 318 97 TYR H H 8.53 0.02 1 319 97 TYR C C 173.08 0.2 1 320 97 TYR CA C 55.83 0.2 1 321 97 TYR CB C 38.41 0.2 1 322 97 TYR N N 120.80 0.15 1 323 98 SER H H 9.44 0.02 1 324 98 SER C C 175.71 0.2 1 325 98 SER CA C 56.79 0.2 1 326 98 SER CB C 67.06 0.2 1 327 98 SER N N 114.46 0.15 1 328 99 GLN H H 9.10 0.02 1 329 99 GLN C C 178.06 0.2 1 330 99 GLN CA C 58.82 0.2 1 331 99 GLN CB C 27.89 0.2 1 332 99 GLN N N 121.95 0.15 1 333 100 ASN H H 8.24 0.02 1 334 100 ASN C C 175.67 0.2 1 335 100 ASN CA C 54.82 0.2 1 336 100 ASN CB C 37.97 0.2 1 337 100 ASN N N 115.25 0.15 1 338 101 ASP H H 7.91 0.02 1 339 101 ASP C C 175.62 0.2 1 342 101 ASP N N 116.62 0.15 1 343 102 LEU H H 7.13 0.02 1 344 102 LEU C C 178.13 0.2 1 347 102 LEU N N 116.52 0.15 1 348 103 VAL H H 7.63 0.02 1 349 103 VAL C C 176.60 0.2 1 350 103 VAL CA C 58.70 0.2 1 352 103 VAL N N 117.07 0.15 1 353 104 GLU H H 8.62 0.02 1 354 104 GLU C C 175.07 0.2 1 355 104 GLU CA C 57.85 0.2 1 357 104 GLU N N 123.79 0.15 1 358 105 TRP H H 8.79 0.02 1 362 105 TRP N N 121.59 0.15 1 363 106 SER H H 8.27 0.02 1 365 107 PRO C C 177.64 0.2 1 366 108 VAL H H 8.51 0.02 1 367 108 VAL C C 179.11 0.2 1 369 108 VAL N N 117.98 0.15 1 370 109 THR H H 9.63 0.02 1 371 109 THR C C 182.02 0.2 1 373 109 THR CB C 69.04 0.2 1 374 109 THR N N 113.65 0.15 1 375 110 GLU H H 8.21 0.02 1 376 110 GLU C C 176.52 0.2 1 377 110 GLU CA C 58.64 0.2 1 378 110 GLU CB C 28.31 0.2 1 379 110 GLU N N 119.88 0.15 1 380 111 LYS H H 7.46 0.02 1 381 111 LYS C C 177.31 0.2 1 382 111 LYS CA C 55.76 0.2 1 384 111 LYS N N 116.20 0.15 1 385 112 HIS H H 7.14 0.02 1 386 112 HIS C C 174.58 0.2 1 387 112 HIS CA C 54.96 0.2 1 388 112 HIS CB C 28.36 0.2 1 389 112 HIS N N 116.18 0.15 1 390 113 LEU H H 7.87 0.02 1 391 113 LEU C C 179.25 0.2 1 394 113 LEU N N 120.14 0.15 1 395 114 THR H H 8.33 0.02 1 396 114 THR C C 175.66 0.2 1 397 114 THR CA C 64.13 0.2 1 399 114 THR N N 109.34 0.15 1 400 115 ASP H H 8.82 0.02 1 401 115 ASP C C 177.72 0.2 1 402 115 ASP CA C 53.76 0.2 1 403 115 ASP CB C 40.28 0.2 1 404 115 ASP N N 117.08 0.15 1 405 116 GLY H H 7.92 0.02 1 406 116 GLY C C 171.67 0.2 1 407 116 GLY CA C 44.77 0.2 1 408 116 GLY N N 110.65 0.15 1 409 117 MET H H 8.33 0.02 1 410 117 MET C C 175.18 0.2 1 411 117 MET CA C 54.73 0.2 1 413 117 MET N N 113.42 0.15 1 414 118 THR H H 8.93 0.02 1 418 118 THR N N 112.14 0.15 1 419 119 VAL H H 8.12 0.02 1 420 119 VAL C C 177.99 0.2 1 421 119 VAL CA C 67.90 0.2 1 423 120 ARG H H 9.09 0.02 1 424 120 ARG C C 177.59 0.2 1 425 120 ARG CA C 59.96 0.2 1 426 120 ARG CB C 31.07 0.2 1 427 120 ARG N N 118.31 0.15 1 428 121 GLU H H 7.64 0.02 1 429 121 GLU C C 180.61 0.2 1 430 121 GLU CA C 58.20 0.2 1 431 121 GLU CB C 29.79 0.2 1 432 121 GLU N N 118.34 0.15 1 433 122 LEU H H 9.13 0.02 1 434 122 LEU C C 179.12 0.2 1 435 122 LEU CA C 57.75 0.2 1 436 122 LEU CB C 40.84 0.2 1 437 122 LEU N N 122.66 0.15 1 438 123 CYS H H 8.17 0.02 1 439 123 CYS C C 176.01 0.2 1 440 123 CYS CA C 62.09 0.2 1 442 123 CYS N N 118.95 0.15 1 443 124 SER H H 7.72 0.02 1 444 124 SER C C 179.07 0.2 1 445 124 SER CA C 61.16 0.2 1 446 124 SER CB C 62.30 0.2 1 447 124 SER N N 113.12 0.15 1 448 125 ALA H H 8.49 0.02 1 449 125 ALA C C 180.31 0.2 1 450 125 ALA CA C 55.41 0.2 1 451 125 ALA CB C 17.33 0.2 1 452 125 ALA N N 124.54 0.15 1 453 126 ALA H H 8.70 0.02 1 454 126 ALA C C 178.45 0.2 1 455 126 ALA CA C 54.80 0.2 1 456 126 ALA CB C 17.25 0.2 1 457 126 ALA N N 120.93 0.15 1 458 127 ILE H H 8.19 0.02 1 459 127 ILE C C 176.59 0.2 1 460 127 ILE N N 113.42 0.15 1 461 128 THR H H 8.96 0.02 1 462 128 THR C C 176.07 0.2 1 463 128 THR N N 109.14 0.15 1 464 129 MET H H 6.87 0.02 1 465 129 MET C C 176.06 0.2 1 466 129 MET CA C 52.75 0.2 1 467 129 MET N N 114.58 0.15 1 468 130 SER H H 7.20 0.02 1 469 130 SER C C 174.49 0.2 1 470 130 SER CA C 58.67 0.2 1 471 130 SER N N 114.84 0.15 1 472 131 ASP H H 7.46 0.02 1 473 131 ASP N N 120.25 0.15 1 474 132 ASN C C 178.14 0.2 1 475 132 ASN CA C 56.08 0.2 1 476 132 ASN CB C 38.91 0.2 1 477 133 THR H H 7.63 0.02 1 478 133 THR C C 176.73 0.2 1 479 133 THR CA C 67.69 0.2 1 480 133 THR CB C 66.87 0.2 1 481 133 THR N N 120.13 0.15 1 482 134 ALA H H 8.90 0.02 1 483 134 ALA C C 178.52 0.2 1 485 134 ALA N N 122.57 0.15 1 486 135 ALA H H 7.22 0.02 1 487 135 ALA C C 178.23 0.2 1 488 135 ALA CB C 19.18 0.2 1 489 135 ALA N N 115.42 0.15 1 490 136 ASN H H 8.03 0.02 1 491 136 ASN C C 178.84 0.2 1 492 136 ASN CA C 55.92 0.2 1 493 136 ASN CB C 37.00 0.2 1 494 136 ASN N N 119.58 0.15 1 495 137 LEU H H 9.08 0.02 1 496 137 LEU C C 181.24 0.2 1 497 137 LEU CA C 58.01 0.2 1 498 137 LEU N N 121.58 0.15 1 499 138 LEU H H 8.20 0.02 1 500 138 LEU C C 180.81 0.2 1 501 138 LEU CA C 57.23 0.2 1 502 138 LEU CB C 42.52 0.2 1 503 138 LEU N N 119.42 0.15 1 504 139 LEU H H 9.21 0.02 1 505 139 LEU C C 180.14 0.2 1 506 139 LEU N N 124.04 0.15 1 507 140 THR H H 8.71 0.02 1 510 140 THR N N 118.06 0.15 1 511 141 THR H H 7.67 0.02 1 512 141 THR C C 175.81 0.2 1 513 141 THR CA C 65.18 0.2 1 514 141 THR CB C 68.73 0.2 1 516 142 ILE H H 6.92 0.02 1 517 142 ILE C C 175.49 0.2 1 518 142 ILE CA C 60.80 0.2 1 519 142 ILE CB C 38.52 0.2 1 520 142 ILE N N 110.87 0.15 1 521 143 GLY H H 7.51 0.02 1 522 143 GLY C C 175.20 0.2 1 523 143 GLY CA C 44.74 0.2 1 524 143 GLY N N 106.38 0.15 1 525 144 GLY H H 8.23 0.02 1 526 144 GLY N N 108.21 0.15 1 527 145 PRO C C 177.94 0.2 1 528 145 PRO CA C 66.01 0.2 1 529 145 PRO CB C 31.71 0.2 1 530 146 LYS H H 8.68 0.02 1 531 146 LYS C C 179.88 0.2 1 532 146 LYS CA C 59.22 0.2 1 533 146 LYS CB C 31.68 0.2 1 534 146 LYS N N 116.02 0.15 1 535 147 GLU H H 7.37 0.02 1 539 147 GLU N N 118.53 0.15 1 540 148 LEU H H 7.67 0.02 1 541 148 LEU C C 178.18 0.2 1 542 148 LEU CA C 57.25 0.2 1 543 148 LEU CB C 40.06 0.2 1 545 149 THR H H 8.19 0.02 1 546 149 THR C C 175.12 0.2 1 547 149 THR CA C 67.75 0.2 1 548 149 THR N N 117.00 0.15 1 549 150 ALA H H 7.91 0.02 1 550 150 ALA C C 179.55 0.2 1 551 150 ALA CA C 55.52 0.2 1 552 150 ALA CB C 17.76 0.2 1 553 150 ALA N N 123.09 0.15 1 554 151 PHE H H 7.81 0.02 1 555 151 PHE C C 177.76 0.2 1 556 151 PHE CA C 60.84 0.2 1 557 151 PHE CB C 37.99 0.2 1 558 151 PHE N N 120.42 0.15 1 559 152 LEU H H 8.07 0.02 1 560 152 LEU C C 179.12 0.2 1 561 152 LEU CA C 57.43 0.2 1 563 152 LEU N N 121.56 0.15 1 564 153 HIS H H 8.91 0.02 1 565 153 HIS C C 179.89 0.2 1 566 153 HIS CA C 59.06 0.2 1 567 153 HIS CB C 28.80 0.2 1 568 153 HIS N N 120.72 0.15 1 569 154 ASN H H 8.39 0.02 1 570 154 ASN C C 176.53 0.2 1 571 154 ASN CA C 55.36 0.2 1 573 154 ASN N N 118.17 0.15 1 574 155 MET H H 7.52 0.02 1 575 155 MET C C 175.88 0.2 1 576 155 MET CA C 55.18 0.2 1 577 155 MET CB C 31.71 0.2 1 578 155 MET N N 115.48 0.15 1 579 156 GLY H H 7.75 0.02 1 582 156 GLY N N 107.68 0.15 1 583 157 ASP H H 8.02 0.02 1 584 157 ASP C C 174.77 0.2 1 585 157 ASP CA C 52.04 0.2 1 586 157 ASP CB C 39.49 0.2 1 588 158 HIS H H 8.34 0.02 1 589 158 HIS C C 175.07 0.2 1 590 158 HIS CA C 54.27 0.2 1 591 158 HIS CB C 29.28 0.2 1 592 158 HIS N N 121.19 0.15 1 593 159 VAL H H 8.82 0.02 1 594 159 VAL C C 176.07 0.2 1 595 159 VAL CA C 63.56 0.2 1 596 159 VAL CB C 34.61 0.2 1 597 159 VAL N N 121.30 0.15 1 598 160 THR H H 9.39 0.02 1 599 160 THR C C 172.84 0.2 1 600 160 THR CA C 64.59 0.2 1 601 160 THR N N 125.61 0.15 1 602 161 ARG H H 8.48 0.02 1 603 161 ARG C C 171.69 0.2 1 604 161 ARG CA C 53.85 0.2 1 605 161 ARG CB C 32.33 0.2 1 606 161 ARG N N 119.70 0.15 1 607 162 LEU H H 7.75 0.02 1 608 162 LEU C C 174.34 0.2 1 609 162 LEU CA C 52.92 0.2 1 610 162 LEU CB C 45.70 0.2 1 611 162 LEU N N 121.06 0.15 1 612 163 ASP H H 10.03 0.02 1 613 163 ASP C C 175.71 0.2 1 614 163 ASP CA C 55.23 0.2 1 616 163 ASP N N 126.71 0.15 1 617 164 ARG H H 9.24 0.02 1 618 164 ARG C C 170.57 0.2 1 619 164 ARG CA C 55.14 0.2 1 620 164 ARG CB C 34.32 0.2 1 621 164 ARG N N 125.16 0.15 1 622 165 TRP H H 7.71 0.02 1 623 165 TRP C C 177.95 0.2 1 624 165 TRP CA C 54.85 0.2 1 625 165 TRP CB C 28.84 0.2 1 626 165 TRP N N 114.55 0.15 1 627 166 GLU H H 9.51 0.02 1 628 166 GLU N N 117.64 0.15 1 629 167 PRO C C 177.90 0.2 1 630 167 PRO CA C 63.59 0.2 1 631 167 PRO CB C 33.98 0.2 1 632 168 GLU H H 8.67 0.02 1 633 168 GLU C C 176.71 0.2 1 634 168 GLU CA C 60.60 0.2 1 635 168 GLU CB C 29.21 0.2 1 636 168 GLU N N 128.28 0.15 1 637 169 LEU H H 7.72 0.02 1 638 169 LEU C C 176.87 0.2 1 639 169 LEU CA C 56.95 0.2 1 640 169 LEU CB C 40.43 0.2 1 641 169 LEU N N 113.05 0.15 1 642 170 ASN H H 7.58 0.02 1 643 170 ASN C C 174.82 0.2 1 644 170 ASN CA C 53.73 0.2 1 646 170 ASN N N 119.87 0.15 1 647 171 GLU H H 7.46 0.02 1 648 171 GLU C C 175.77 0.2 1 649 171 GLU CA C 60.54 0.2 1 650 171 GLU CB C 30.68 0.2 1 651 171 GLU N N 120.67 0.15 1 652 172 ALA H H 9.51 0.02 1 653 172 ALA C C 176.56 0.2 1 655 172 ALA CB C 17.79 0.2 1 656 172 ALA N N 118.23 0.15 1 657 173 ILE H H 8.67 0.02 1 658 173 ILE N N 120.75 0.15 1 659 174 PRO C C 177.56 0.2 1 660 174 PRO CA C 64.06 0.2 1 661 174 PRO CB C 31.24 0.2 1 662 175 ASN H H 8.91 0.02 1 663 175 ASN C C 173.30 0.2 1 664 175 ASN CA C 54.99 0.2 1 665 175 ASN CB C 37.20 0.2 1 666 175 ASN N N 115.82 0.15 1 667 176 ASP H H 7.61 0.02 1 668 176 ASP C C 177.37 0.2 1 669 176 ASP CA C 53.03 0.2 1 670 176 ASP CB C 41.82 0.2 1 671 176 ASP N N 120.76 0.15 1 672 177 GLU H H 9.00 0.02 1 673 177 GLU C C 177.66 0.2 1 674 177 GLU CA C 56.66 0.2 1 676 177 GLU N N 126.37 0.15 1 677 178 ARG H H 8.12 0.02 1 678 178 ARG C C 177.22 0.2 1 679 178 ARG CA C 57.13 0.2 1 680 178 ARG N N 122.25 0.15 1 681 179 ASP H H 8.81 0.02 1 682 179 ASP C C 177.98 0.2 1 683 179 ASP CA C 55.67 0.2 1 684 179 ASP CB C 42.25 0.2 1 685 179 ASP N N 115.51 0.15 1 686 180 THR H H 7.29 0.02 1 687 180 THR C C 173.15 0.2 1 688 180 THR CA C 59.59 0.2 1 689 180 THR N N 104.73 0.15 1 690 181 THR H H 8.24 0.02 1 691 181 THR C C 173.63 0.2 1 692 181 THR CA C 59.55 0.2 1 693 181 THR CB C 68.81 0.2 1 694 181 THR N N 111.69 0.15 1 695 182 MET H H 8.54 0.02 1 696 182 MET N N 117.82 0.15 1 697 183 PRO C C 176.93 0.2 1 698 183 PRO CA C 67.23 0.2 1 699 183 PRO CB C 32.27 0.2 1 700 184 ALA H H 8.70 0.02 1 701 184 ALA C C 179.13 0.2 1 702 184 ALA CA C 55.92 0.2 1 703 184 ALA CB C 18.25 0.2 1 704 184 ALA N N 114.70 0.15 1 705 185 ALA H H 6.88 0.02 1 706 185 ALA C C 178.04 0.2 1 707 185 ALA CA C 54.50 0.2 1 709 185 ALA N N 121.02 0.15 1 710 186 MET H H 8.34 0.02 1 711 186 MET C C 177.93 0.2 1 712 186 MET CA C 57.11 0.2 1 713 186 MET CB C 30.83 0.2 1 714 186 MET N N 116.05 0.15 1 715 187 ALA H H 8.29 0.02 1 716 187 ALA C C 178.48 0.2 1 717 187 ALA CA C 55.55 0.2 1 719 187 ALA N N 117.75 0.15 1 720 188 THR H H 7.72 0.02 1 721 188 THR C C 177.42 0.2 1 722 188 THR CA C 66.33 0.2 1 723 188 THR CB C 68.70 0.2 1 724 188 THR N N 112.23 0.15 1 725 189 THR H H 9.07 0.02 1 726 189 THR C C 175.81 0.2 1 727 189 THR CA C 66.93 0.2 1 728 189 THR N N 124.55 0.15 1 729 190 LEU H H 8.87 0.02 1 730 190 LEU C C 178.28 0.2 1 731 190 LEU CA C 57.82 0.2 1 733 190 LEU N N 121.29 0.15 1 734 191 ARG H H 8.15 0.02 1 735 191 ARG C C 178.80 0.2 1 737 191 ARG CB C 28.73 0.2 1 738 191 ARG N N 118.28 0.15 1 739 192 LYS H H 8.12 0.02 1 743 192 LYS N N 120.91 0.15 1 744 193 LEU H H 8.07 0.02 1 745 193 LEU C C 177.52 0.2 1 746 193 LEU CA C 57.70 0.2 1 749 194 LEU H H 8.07 0.02 1 750 194 LEU C C 178.37 0.2 1 751 194 LEU CA C 56.41 0.2 1 753 194 LEU N N 111.39 0.15 1 754 195 THR H H 7.99 0.02 1 755 195 THR C C 174.71 0.2 1 756 195 THR CA C 60.72 0.2 1 758 195 THR N N 105.32 0.15 1 759 196 GLY H H 7.91 0.02 1 760 196 GLY C C 174.08 0.2 1 761 196 GLY CA C 44.76 0.2 1 762 196 GLY N N 110.47 0.15 1 763 197 GLU H H 8.40 0.02 1 764 197 GLU C C 176.48 0.2 1 765 197 GLU CA C 55.77 0.2 1 766 197 GLU CB C 29.32 0.2 1 767 197 GLU N N 116.70 0.15 1 768 198 LEU H H 7.49 0.02 1 769 198 LEU C C 177.48 0.2 1 770 198 LEU CB C 42.48 0.2 1 771 198 LEU N N 122.60 0.15 1 772 199 LEU H H 8.71 0.02 1 776 199 LEU N N 118.20 0.15 1 777 200 THR H H 8.98 0.02 1 778 200 THR C C 175.43 0.2 1 779 200 THR CA C 61.94 0.2 1 780 200 THR CB C 71.57 0.2 1 782 201 LEU H H 8.71 0.02 1 783 201 LEU C C 179.13 0.2 1 784 201 LEU CA C 58.68 0.2 1 785 201 LEU CB C 41.35 0.2 1 786 201 LEU N N 122.97 0.15 1 787 202 ALA H H 8.47 0.02 1 788 202 ALA C C 181.55 0.2 1 789 202 ALA CA C 54.78 0.2 1 791 202 ALA N N 118.63 0.15 1 792 203 SER H H 7.76 0.02 1 793 203 SER C C 175.12 0.2 1 794 203 SER CA C 62.61 0.2 1 795 203 SER N N 118.40 0.15 1 796 204 ARG H H 9.08 0.02 1 797 204 ARG C C 178.59 0.2 1 798 204 ARG CA C 60.16 0.2 1 799 204 ARG CB C 30.76 0.2 1 800 204 ARG N N 123.51 0.15 1 801 205 GLN H H 8.24 0.02 1 802 205 GLN C C 177.17 0.2 1 803 205 GLN CA C 57.36 0.2 1 805 205 GLN N N 117.54 0.15 1 806 206 GLN H H 7.62 0.02 1 807 206 GLN C C 177.92 0.2 1 808 206 GLN CA C 56.77 0.2 1 809 206 GLN CB C 28.36 0.2 1 810 206 GLN N N 118.04 0.15 1 811 207 LEU H H 7.99 0.02 1 812 207 LEU C C 178.33 0.2 1 815 207 LEU N N 120.97 0.15 1 816 208 ILE H H 7.81 0.02 1 817 208 ILE C C 177.31 0.2 1 820 208 ILE N N 118.08 0.15 1 821 209 ASP H H 8.69 0.02 1 822 209 ASP C C 181.49 0.2 1 825 209 ASP N N 122.31 0.15 1 826 210 TRP H H 8.06 0.02 1 830 210 TRP N N 120.43 0.15 1 831 211 MET H H 7.76 0.02 1 832 211 MET C C 180.32 0.2 1 835 212 GLU H H 9.64 0.02 1 836 212 GLU C C 178.17 0.2 1 837 212 GLU CA C 59.46 0.2 1 839 212 GLU N N 124.59 0.15 1 840 213 ALA H H 7.35 0.02 1 841 213 ALA C C 175.56 0.2 1 842 213 ALA CA C 51.49 0.2 1 843 213 ALA N N 119.65 0.15 1 844 214 ASP H H 7.01 0.02 1 845 214 ASP C C 176.78 0.2 1 846 214 ASP CA C 56.04 0.2 1 847 214 ASP CB C 43.85 0.2 1 848 214 ASP N N 115.83 0.15 1 849 215 LYS H H 9.22 0.02 1 850 215 LYS C C 179.68 0.2 1 851 215 LYS CA C 56.46 0.2 1 852 215 LYS N N 128.01 0.15 1 853 216 VAL H H 7.27 0.02 1 854 216 VAL C C 175.01 0.2 1 855 216 VAL CA C 60.52 0.2 1 856 216 VAL N N 110.22 0.15 1 857 217 ALA H H 8.89 0.02 1 858 217 ALA C C 178.40 0.2 1 859 217 ALA CB C 20.59 0.2 1 860 217 ALA N N 125.34 0.15 1 861 218 GLY H H 8.94 0.02 1 862 218 GLY N N 114.22 0.15 1 863 219 PRO C C 175.22 0.2 1 864 219 PRO CA C 63.81 0.2 1 865 220 LEU H H 6.82 0.02 1 866 220 LEU C C 176.47 0.2 1 867 220 LEU CA C 51.68 0.2 1 868 220 LEU N N 113.95 0.15 1 869 221 LEU H H 6.90 0.02 1 870 221 LEU C C 178.60 0.2 1 871 221 LEU CA C 57.79 0.2 1 872 221 LEU N N 123.35 0.15 1 873 222 ARG H H 8.97 0.02 1 874 222 ARG C C 177.53 0.2 1 875 222 ARG CA C 59.66 0.2 1 876 222 ARG CB C 29.76 0.2 1 877 222 ARG N N 115.99 0.15 1 878 223 SER H H 7.04 0.02 1 879 223 SER C C 174.09 0.2 1 880 223 SER CA C 60.74 0.2 1 881 223 SER CB C 63.09 0.2 1 882 223 SER N N 113.33 0.15 1 883 224 ALA H H 7.29 0.02 1 887 224 ALA N N 122.81 0.15 1 888 225 LEU H H 6.72 0.02 1 890 226 PRO C C 175.86 0.2 1 891 226 PRO CA C 61.18 0.2 1 892 226 PRO CB C 31.75 0.2 1 893 227 ALA H H 8.26 0.02 1 894 227 ALA C C 179.48 0.2 1 895 227 ALA CA C 53.25 0.2 1 897 227 ALA N N 122.70 0.15 1 898 228 GLY H H 8.84 0.02 1 901 228 GLY N N 110.10 0.15 1 902 229 TRP H H 7.64 0.02 1 903 229 TRP C C 175.43 0.2 1 904 229 TRP CA C 58.81 0.2 1 906 230 PHE H H 9.39 0.02 1 907 230 PHE C C 174.86 0.2 1 908 230 PHE CA C 56.58 0.2 1 909 230 PHE CB C 41.90 0.2 1 910 230 PHE N N 122.91 0.15 1 911 231 ILE H H 7.53 0.02 1 912 231 ILE C C 169.55 0.2 1 913 231 ILE CA C 60.25 0.2 1 914 231 ILE N N 123.34 0.15 1 915 232 ALA H H 8.47 0.02 1 916 232 ALA C C 174.97 0.2 1 917 232 ALA CA C 50.45 0.2 1 918 232 ALA CB C 21.20 0.2 1 919 232 ALA N N 128.04 0.15 1 920 233 ASP H H 8.26 0.02 1 921 233 ASP C C 172.75 0.2 1 922 233 ASP N N 123.94 0.15 1 923 234 LYS H H 8.05 0.02 1 924 234 LYS C C 178.37 0.2 1 925 234 LYS N N 109.55 0.15 1 926 235 SER H H 8.28 0.02 1 927 235 SER C C 174.09 0.2 1 928 235 SER N N 115.49 0.15 1 929 236 GLY H H 8.59 0.02 1 930 236 GLY N N 102.88 0.15 1 931 237 ALA C C 175.48 0.2 1 932 238 GLY H H 7.72 0.02 1 933 238 GLY C C 172.79 0.2 1 934 238 GLY CA C 45.22 0.2 1 935 238 GLY N N 107.35 0.15 1 936 239 GLU H H 7.87 0.02 1 937 239 GLU C C 176.05 0.2 1 938 239 GLU CA C 55.93 0.2 1 940 239 GLU N N 117.03 0.15 1 941 240 ARG H H 9.48 0.02 1 942 240 ARG C C 176.11 0.2 1 943 240 ARG CA C 57.10 0.2 1 945 240 ARG N N 118.06 0.15 1 946 241 GLY H H 8.80 0.02 1 947 241 GLY C C 177.37 0.2 1 948 241 GLY CA C 46.26 0.2 1 949 241 GLY N N 102.83 0.15 1 950 242 SER H H 7.54 0.02 1 951 242 SER C C 174.98 0.2 1 952 242 SER CA C 59.79 0.2 1 953 242 SER CB C 64.13 0.2 1 954 242 SER N N 119.23 0.15 1 955 243 ARG H H 9.06 0.02 1 956 243 ARG C C 174.27 0.2 1 957 243 ARG CA C 54.22 0.2 1 959 243 ARG N N 126.22 0.15 1 960 244 GLY H H 8.52 0.02 1 961 244 GLY C C 170.35 0.2 1 962 244 GLY CA C 46.52 0.2 1 963 244 GLY N N 111.61 0.15 1 964 245 ILE H H 9.18 0.02 1 965 245 ILE C C 170.51 0.2 1 966 245 ILE CA C 59.42 0.2 1 967 245 ILE N N 116.74 0.15 1 968 246 ILE H H 8.13 0.02 1 969 246 ILE C C 174.81 0.2 1 970 246 ILE CA C 58.23 0.2 1 971 246 ILE CB C 39.85 0.2 1 972 246 ILE N N 118.02 0.15 1 973 247 ALA H H 9.01 0.02 1 974 247 ALA C C 174.67 0.2 1 975 247 ALA CA C 51.12 0.2 1 976 247 ALA CB C 24.25 0.2 1 977 247 ALA N N 122.70 0.15 1 978 248 ALA H H 9.36 0.02 1 979 248 ALA C C 175.66 0.2 1 980 248 ALA CA C 50.08 0.2 1 981 248 ALA CB C 20.17 0.2 1 982 248 ALA N N 122.83 0.15 1 983 249 LEU H H 9.34 0.02 1 984 249 LEU C C 176.75 0.2 1 985 249 LEU CA C 54.85 0.2 1 986 249 LEU CB C 46.34 0.2 1 987 249 LEU N N 121.17 0.15 1 988 250 GLY H H 8.83 0.02 1 989 250 GLY N N 110.44 0.15 1 990 251 PRO C C 176.08 0.2 1 991 251 PRO CA C 60.59 0.2 1 992 251 PRO CB C 31.34 0.2 1 993 252 ASP H H 7.95 0.02 1 994 252 ASP C C 177.20 0.2 1 995 252 ASP CA C 54.74 0.2 1 996 252 ASP CB C 38.99 0.2 1 997 252 ASP N N 114.30 0.15 1 998 253 GLY H H 7.73 0.02 1 999 253 GLY C C 173.45 0.2 1 1000 253 GLY CA C 46.64 0.2 1 1001 253 GLY N N 100.17 0.15 1 1002 254 LYS H H 7.20 0.02 1 1003 254 LYS N N 119.33 0.15 1 1004 255 PRO C C 176.14 0.2 1 1005 255 PRO CA C 61.94 0.2 1 1007 256 SER H H 8.56 0.02 1 1008 256 SER C C 174.58 0.2 1 1009 256 SER CA C 58.76 0.2 1 1010 256 SER CB C 65.48 0.2 1 1011 256 SER N N 112.48 0.15 1 1012 257 ARG H H 8.85 0.02 1 1013 257 ARG C C 174.58 0.2 1 1014 257 ARG CA C 55.68 0.2 1 1015 257 ARG CB C 35.93 0.2 1 1016 257 ARG N N 123.06 0.15 1 1017 258 ILE H H 9.26 0.02 1 1018 258 ILE C C 174.62 0.2 1 1019 258 ILE CA C 58.58 0.2 1 1020 258 ILE CB C 40.46 0.2 1 1021 258 ILE N N 122.63 0.15 1 1022 259 VAL H H 9.23 0.02 1 1023 259 VAL C C 173.60 0.2 1 1024 259 VAL CA C 59.33 0.2 1 1025 259 VAL CB C 35.99 0.2 1 1026 259 VAL N N 126.22 0.15 1 1027 260 VAL H H 8.26 0.02 1 1028 260 VAL C C 173.85 0.2 1 1029 260 VAL CA C 60.31 0.2 1 1030 260 VAL CB C 35.54 0.2 1 1031 260 VAL N N 125.30 0.15 1 1032 261 ILE H H 8.36 0.02 1 1033 261 ILE C C 174.38 0.2 1 1034 261 ILE CA C 59.81 0.2 1 1035 261 ILE CB C 41.53 0.2 1 1036 261 ILE N N 123.07 0.15 1 1037 262 TYR H H 8.75 0.02 1 1041 262 TYR N N 122.22 0.15 1 1042 263 THR H H 8.96 0.02 1 1047 264 THR H H 8.04 0.02 1 1051 265 GLY H H 8.69 0.02 1 1055 266 SER H H 8.36 0.02 1 1059 267 GLN H H 8.84 0.02 1 1064 268 ALA H H 8.71 0.02 1 1069 269 THR H H 8.66 0.02 1 1073 270 MET H H 9.14 0.02 1 1078 271 ASP H H 8.46 0.02 1 1083 272 GLU H H 7.64 0.02 1 1088 273 ARG H H 8.13 0.02 1 1093 274 ASN H H 8.78 0.02 1 1098 275 ARG H H 8.04 0.02 1 1103 276 GLN H H 7.81 0.02 1 1108 277 ILE H H 7.43 0.02 1 1113 278 ALA H H 8.72 0.02 1 1118 279 GLU H H 8.45 0.02 1 1123 280 ILE H H 8.05 0.02 1 1127 281 GLY H H 8.37 0.02 1 1131 282 ALA H H 8.65 0.02 1 1136 283 SER H H 7.52 0.02 1 1141 284 LEU H H 8.23 0.02 1 1146 285 ILE H H 7.98 0.02 1 1151 286 LYS H H 8.13 0.02 1 1156 287 HIS H H 7.64 0.02 1 1161 288 TRP H H 7.27 0.02 1 stop_ save_