HtrA1 bound to an optimized peptide: NMR assignment of PDZ domain and ligand resonances
GSHMKKYIGI RMMSLTSSKA KELKDRHRDF PDVISGAYII EVIPDTPAEA GGLKENDVII SINGQSVVSA NDVSDVIKRE STLNMVVRRG NEDIMITVIP EEIDP
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 89.9 % (1158 of 1288) | 92.7 % (621 of 670) | 86.3 % (434 of 503) | 89.6 % (103 of 115) |
Backbone | 91.1 % (603 of 662) | 94.2 % (213 of 226) | 89.1 % (293 of 329) | 90.7 % (97 of 107) |
Sidechain | 89.1 % (651 of 731) | 91.9 % (408 of 444) | 84.9 % (237 of 279) | 75.0 % (6 of 8) |
Aromatic | 29.3 % (17 of 58) | 48.3 % (14 of 29) | 11.1 % (3 of 27) | 0.0 % (0 of 2) |
Methyl | 97.1 % (134 of 138) | 100.0 % (69 of 69) | 94.2 % (65 of 69) |
1. HtrA1-PDZ
GSHMKKYIGI RMMSLTSSKA KELKDRHRDF PDVISGAYII EVIPDTPAEA GGLKENDVII SINGQSVVSA NDVSDVIKRE STLNMVVRRG NEDIMITVIP EEIDP2. synthetic peptide H1-C1
DSRIWWVSolvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0, Details 25mM sodium phosphate pH 6.0 + 1mM sodium azide - slight excess of peptide
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HtrA1-PDZ | [U-15N] | 2 mM | |
2 | synthetic peptide H1-C1 | natural abundance | 4 mM | |
3 | sodium phosphate | natural abundance | 25 mM | |
4 | sodium azide | natural abundance | 1 mM |
Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0, Details 25mM sodium phosphate pH 6.0 + 1mM sodium azide - excess peptide
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
5 | HtrA1-PDZ | [U-13C; U-15N] | 2 mM | |
6 | synthetic peptide H1-C1 | natural abundance | 4 mM | |
7 | sodium phosphate | natural abundance | 25 mM | |
8 | sodium azide | natural abundance | 1 mM |
Solvent system 100% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0, Details 25mM sodium phosphate pH 6.0 + 1mM sodium azide - excess of peptide
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
9 | HtrA1-PDZ | [U-13C; U-15N] | 2 mM | |
10 | synthetic peptide H1-C1 | natural abundance | 4 mM | |
11 | sodium phosphate | natural abundance | 25 mM | |
12 | sodium azide | natural abundance | 1 mM |
Solvent system 100% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0, Details 25mM sodium phosphate pH 6.0 + 1mM sodium azide -slight excess of peptide; partial carbon label for stereo specific methyl assignment
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
13 | HtrA1-PDZ | [U-10% 13C; U-99% 15N] | 2 mM | |
14 | synthetic peptide H1-C1 | natural abundance | 4 mM | |
15 | sodium phosphate | natural abundance | 25 mM | |
16 | sodium azide | natural abundance | 1 mM |
Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0, Details 25mM sodium phosphate pH 6.0 + 1mM sodium azide - slight excess of PDZ
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
17 | HtrA1-PDZ | [U-13C; U-15N] | 2 mM | |
18 | synthetic peptide H1-C1 | natural abundance | 1.8 mM | |
19 | sodium phosphate | natural abundance | 25 mM | |
20 | sodium azide | natural abundance | 1 mM |
Solvent system 100% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0, Details 25mM sodium phosphate pH 6.0 + 1mM sodium azide - slight excess of PDZ
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
21 | HtrA1-PDZ | [U-13C; U-15N] | 2 mM | |
22 | synthetic peptide H1-C1 | natural abundance | 1.8 mM | |
23 | sodium phosphate | natural abundance | 25 mM | |
24 | sodium azide | natural abundance | 1 mM |