PDBsum entry 2as9

Hydrolase

PDB id: 2as9

Contents

Protein chains

207 a.a. *

195 a.a. *

Metals

_ZN ×7

Waters ×222

* Residue conservation analysis

PDB id:

2as9

Name:

Hydrolase

Title:

Functional and structural characterization of spl proteases from staphylococcus aureus

Structure:

Serine protease. Chain: a, b. Fragment: residues 1-210. Synonym: splc serine protease. Engineered: yes

Source:

Staphylococcus aureus. Organism_taxid: 1280. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.

Resolution:

1.70Å R-factor: 0.213 R-free: 0.243

Authors:

G.M.Popowicz,G.Dubin,J.Stec-Niemczyk,A.Czarny,A.Dubin,J.Potempa, T.A.Holak

Key ref:

G.M.Popowicz et al. (2006). Functional and structural characterization of Spl proteases from Staphylococcus aureus. J Mol Biol, 358, 270-279. PubMed id: 16516230 DOI: 10.1016/j.jmb.2006.01.098

Date:

23-Aug-05 Release date: 06-Sep-05

Protein chain

Q2FXC4  (SPLC_STAA8) -  Serine protease SplC from Staphylococcus aureus (strain NCTC 8325 / PS 47)

Seq: 239 a.a.

Struc: 207 a.a.*

Protein chain

Q2FXC4  (SPLC_STAA8) -  Serine protease SplC from Staphylococcus aureus (strain NCTC 8325 / PS 47)

Seq: 239 a.a.

Struc: 195 a.a.*

* PDB and UniProt seqs differ at 3 residue positions (black crosses)

Enzyme reactions

• Enzyme class: Chains A, B: E.C.3.4.21.- - ?????

DOI no: 10.1016/j.jmb.2006.01.098

J Mol Biol 358:270-279 (2006)

PubMed id: 16516230

Functional and structural characterization of Spl proteases from Staphylococcus aureus.

G.M.Popowicz, G.Dubin, J.Stec-Niemczyk, A.Czarny, A.Dubin, J.Potempa, T.A.Holak.

ABSTRACT

Staphylococcus aureus is the major cause of nosocomial infections world-wide, with increasing prevalence of community-acquired diseases. The recent dramatic increase in multi-antibiotic resistance, including resistance to the last-resort drug, vancomycin, together with the lack of an effective vaccine highlight the need for better understanding of S.aureus pathogenicity. Comparative analysis of available bacterial genomes allows for the identification of previously uncharacterized S.aureus genes with potential roles in pathogenicity. A good example is a cluster of six serine protease-like (spl) genes encompassed in one operon, which encode for putative proteases with similarity to staphylococcal glutamylendopeptidase (V8 protease). Here, we describe an efficient expression system for the production of recombinant SplB and SplC proteases in Escherichia coli, together with structural and functional characterization of the purified enzymes. A unique mechanism of cytoplasm protection against activity of misdirected SplB was uncovered. Apparently, the co-translated signal peptide maintains protease latency until it is cleaved by the signal peptidase during protein secretion. Furthermore, the crystal structure of the SplC protease revealed a fold resembling that of the V8 protease and epidermolytic toxins. Arrangement of the active site cleft and substrate-binding pocket of SplC explains the mechanism of enzyme latency and suggests that some Spl proteases possess restricted substrate specificity similar to that of the V8 protease and epidermolytic toxins.

Selected figure(s)

Figure 2.
Figure 2. Surface representation of the SplC molecule. The catalytic triad (yellow) is blocked by Gly175–Gly176 and the whole Tyr174–Tyr184 loop region interferes with enzymatic activity. The specificity region built by Val119, Phe149, Ile153 and Val173 enables binding of a medium-sized hydrophobic residue.

Figure 4.
Figure 4. Structure of SplC (red) superimposed on the V8 protease (blue). (a) An overall view of the whole model showing a high degree of structural similarity. (b) The detailed view of the active site and specificity region. The structures appear to be almost identical, except for the Tyr174–Tyr184 loop, which is β-structured in V8 while flexible in SplC. The “locked” state of the catalytic His40 is evident.

The above figures are reprinted by permission from Elsevier: J Mol Biol (2006, 358, 270-279) copyright 2006.

Figures were selected by an automated process.