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SOLUTION STRUCTURE OF THE S. AUREUS SORTASE A-SUBSTRATE COMPLEX
Authors
Suree, N., Liew, C.KONG., Villareal, V.A., Thieu, W., Fadeev, E.A., Clemens, J.J., Jung, M.E., Clubb, R.T.
Assembly
Sortase A-substrate complex
Entity
1. Sortase A (polymer, Thiol state: all disulfide bound), 148 monomers, 16727.78 Da Detail

MQAKPQIPKD KSKVAGYIEI PDADIKEPVY PGPATPEQLN RGVSFAEENE SLDDQNISIA GHTFIDRPNY QFTNLKAAKK GSMVYFKVGN ETRKYKMTSI RDVKPTDVGV LDEQKGKDKQ LTLITCDDYN EKTGVWEKRK IFVATEVK


2. Cbz-LPAT (polymer, Thiol state: not present), 5 monomers, 555.1289 Da Detail

XLPAX


3. CA (non-polymer), 40.078 Da
Total weight
17322.986 Da
Max. entity weight
16727.78 Da
Entity Connection
disulfide 1 Detail

IDTypeValue orderAtom ID 1Atom ID 2
1disulfidesing1:CYS126:SG2:B275:SG

Source organism
Staphylococcus aureus
Exptl. method
solution NMR
Refine. method
SIMULATED ANNEALING
Data set
assigned_chemical_shifts
Chem. Shift Complete
Sequence coverage: 98.7 %, Completeness: 91.8 %, Completeness (bb): 97.7 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All91.8 % (1648 of 1796)90.8 % (859 of 946)91.5 % (635 of 694)98.7 % (154 of 156)
Backbone97.7 % (866 of 886)99.3 % (299 of 301)96.2 % (427 of 444)99.3 % (140 of 141)
Sidechain87.5 % (921 of 1052)86.8 % (560 of 645)88.5 % (347 of 392)93.3 % (14 of 15)
Aromatic45.6 % (52 of 114)45.6 % (26 of 57)44.6 % (25 of 56)100.0 % (1 of 1)
Methyl100.0 % (152 of 152)100.0 % (76 of 76)100.0 % (76 of 76)

1. Sortase A

MQAKPQIPKD KSKVAGYIEI PDADIKEPVY PGPATPEQLN RGVSFAEENE SLDDQNISIA GHTFIDRPNY QFTNLKAAKK GSMVYFKVGN ETRKYKMTSI RDVKPTDVGV LDEQKGKDKQ LTLITCDDYN EKTGVWEKRK IFVATEVK

2. Cbz-LPAT

XLPAX

Sample #1

Solvent system 93% H2O, 7% D2O, Pressure 1 atm, Temperature 302 K, pH 6.0


#NameIsotope labelingTypeConcentration
1Sortase A[U-100% 13C; U-100% 15N]1 mM
2Cbz-LPAT*natural abundance1 mM
3TRISnatural abundance50 mM
4sodium chloridenatural abundance100 mM
5Calcium Chloridenatural abundance20 mM
6sodium azidenatural abundance0.01 %
7H2Onatural abundance93 %
8D2O[U-100% 2H]7 %
Sample #2

Solvent system 93% H2O, 7% D2O, Pressure 1 atm, Temperature 302 K, pH 6.0


#NameIsotope labelingTypeConcentration
9Sortase A[U-100% 15N]1 mM
10Cbz-LPAT*natural abundance1 mM
11TRISnatural abundance50 mM
12sodium chloridenatural abundance100 mM
13Calcium Chloridenatural abundance20 mM
14sodium azidenatural abundance0.01 %
15H2Onatural abundance93 %
16D2O[U-100% 2H]7 %
Sample #3

Solvent system 93% H2O, 7% D2O, Pressure 1 atm, Temperature 302 K, pH 6.0


#NameIsotope labelingTypeConcentration
17Sortase A[U-100% 13C; U-100% 15N]1 mM
18Cbz-LPAT*natural abundance1 mM
19TRISnatural abundance50 mM
20sodium chloridenatural abundance100 mM
21Calcium Chloridenatural abundance20 mM
22sodium azidenatural abundance0.01 %
23H2Onatural abundance93 %
24D2O[U-100% 2H]7 %

Protein Blocks Logo
Calculated from 20 models in PDB: 2KID, Strand ID: A Detail


Release date
2009-07-15
Citation
The structure of the Staphylococcus aureus sortase-substrate complex reveals how the universally conserved LPXTG sorting signal is recognized
Suree, N., Liew, C.KONG., Villareal, V.A., Thieu, W., Fadeev, E.A., Clemens, J.J., Jung, M.E., Clubb, R.T.
J. Biol. Chem. (2009), 284, 24465-24477, PubMed 19592495 , DOI 10.1074/jbc.M109.022624 ,
Related entities 1. Sortase A, : 1 entities Detail
Experiments performed 14 experiments Detail
NMR combined restraints 9 contents Detail
Keywords Cell Wall, complex, enzyme, NMR, sortase, EIGHT STRANDED BETA BARREL, ENZYME-SUBSTRATE COMPLEX, SOLUTION NMR, SORTASE, TRANSPEPTIDASE