PDBsum entry 2k3a

Hydrolase

PDB id: 2k3a

Contents

Protein chain

155 a.a. *

* Residue conservation analysis

PDB id:

2k3a

Name:

Hydrolase

Title:

Nmr solution structure of staphylococcus saprophyticus chap (cysteine, histidine-dependent amidohydrolases/peptidases) domain protein. Northeast structural genomics consortium target syr11

Structure:

Chap domain protein. Chain: a. Engineered: yes

Source:

Staphylococcus saprophyticus subsp. Saprophyticus atcc 15305. Organism_taxid: 342451. Strain: atcc 15305 / dsm 20229. Gene: ssp0609. Expressed in: escherichia coli. Expression_system_taxid: 562. Other_details: syr11-21.5

NMR struc:

20 models

Authors:

P.Rossi,J.M.Aramini,C.X.Chen,C.Nwosu,K.C.Cunningham,L.A.Owens,R.Xiao, J.Liu,M.C.Baran,G.Swapna,T.B.Acton,B.Rost,G.T.Montelione,Northeast Structural Genomics Consortium (Nesg)

Key ref:

P.Rossi et al. (2009). Structural elucidation of the Cys-His-Glu-Asn proteolytic relay in the secreted CHAP domain enzyme from the human pathogen Staphylococcus saprophyticus. Proteins, 74, 515-519. PubMed id: 18951393 DOI: 10.1002/prot.22267

Date:

29-Apr-08 Release date: 13-May-08

Supersedes:

2jrn

Protein chain

Q49ZM2  (Q49ZM2_STAS1) -  Putative secretory antigen from Staphylococcus saprophyticus subsp. saprophyticus (strain ATCC 15305 / DSM 20229 / NCIMB 8711 / NCTC 7292 / S-41)

Seq: 155 a.a.

Struc: 155 a.a.

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1002/prot.22267

Proteins 74:515-519 (2009)

PubMed id: 18951393

Structural elucidation of the Cys-His-Glu-Asn proteolytic relay in the secreted CHAP domain enzyme from the human pathogen Staphylococcus saprophyticus.

P.Rossi, J.M.Aramini, R.Xiao, C.X.Chen, C.Nwosu, L.A.Owens, M.Maglaqui, R.Nair, M.Fischer, T.B.Acton, B.Honig, B.Rost, G.T.Montelione.

ABSTRACT

No abstract given.

Selected figure(s)

Figure 1.
Figure 1. (A) Stereoview of the ribbon representation of the lowest energy conformer (lowest CNS energy) from the ensemble of deposited solution NMR structures of full length SSP0609 (PDB_ID: 2K3A). Only residues 50-155 are shown. The secondary structural elements are labeled and the triad sidechain residues (Cys57, His109, Glu126) are highlighted as red sticks. (B) ConSurf images of SSP0609 for identical size and orientation as (A) ConSurf analysis was conducted for the entire CHAP protein domain family, standard ConSurf residue coloring reflecting the degree of residue conservation over the entire family were used (color scheme: magenta, highly conserved; cyan, variable). (C) Electrostatic potential surface diagrams for SSP0609 shown in the same orientation as (A) and (B).

Figure 2.
Figure 2. (A) Characterization of His109 and His153 tautomer state by 2D ^1H-^15N HMQC spectrum of [U-^15N, 5%^13C] SSP0609 at pH 6.5 and 25°C, obtained on an 800 MHz NMR spectrometer. Acquisition parameters: ^15N carrier at 205 ppm, ^2J(^15N-^1H) = 22Hz, 2 sec relaxation delay, acquisition dimensions 2048 × 200 complex points, sweep widths 20 ppm (^1H) × 200 ppm (^15N), and 128 transients per t[1] increment. (B) Plots of histidine H^ 1 chemical shift versus pH for U-^13C,^15N S. saprophyticus SSP0609 at 298 K, obtained by 2D ^1H-^13C HSQC NMR. Red, His109; green, His153 (surface exposed); blue, C-terminal His tag (control); pK[a] values for each are indicated in the plot. (C) Detailed view of SSP0609 active site showing the cysteine peptidase catalytic triad formed by Cys57, His109, Glu126, and the additional participating group Asn128. Showing here are the strongly conserved Gln56, Thr58, and Gly74 in the active site. Key hydrogen bond distances: Cys57 S^ -His109 N^ 1 = 3.3 Å, His109 N^ 2-Glu126 O^ 2 = 3.2 Å, Glu126 O^ 1-Asn128 N^ 2 = 4.1 Å.

The above figures are reprinted by permission from John Wiley & Sons, Inc.: Proteins (2009, 74, 515-519) copyright 2009.