PDBsum entry: 1go7

Hydrolase

PDB id: 1go7

Contents

Protein chain

462 a.a. *

Ligands

PO4

Metals

_CA ×7

_ZN

Waters ×335

* Residue conservation analysis

PDB id:

1go7

Name:

Hydrolase

Title:

The metzincin's methionine: prtc m226c-e189k double mutant

Structure:

ProteasE C. Chain: p. Engineered: yes. Mutation: yes

Source:

Erwinia chrysanthemi. Organism_taxid: 556. Expressed in: escherichia coli. Expression_system_taxid: 562.

Biol. unit:

Dimer (from PDB file)

Resolution:

2.1Å R-factor: 0.172 R-free: 0.204

Authors:

T.Hege

Key ref:

T.Hege et al. (2001). Crystal structure of a complex between Pseudomonas aeruginosa alkaline protease and its cognate inhibitor: inhibition by a zinc-NH2 coordinative bond. J Biol Chem, 276, 35087-35092. PubMed id: 11445573 DOI: 10.1074/jbc.M104020200

Date:

20-Oct-01 Release date: 17-Oct-02

Protein chain

P16317  (PRTC_ERWCH) -  Serralysin C

Seq: 479 a.a.

Struc: 462 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.4.24.40 - Serralysin.
• Reaction: Preferential cleavage of bonds with hydrophobic residues in P1'.
• Cofactor: Zinc

 Gene Ontology (GO) functional annotation 

• Cellular component: extracellular region (3 terms)
• Biological process: proteolysis (1 term)
• Biochemical function: hydrolase activity (7 terms)

DOI no: 10.1074/jbc.M104020200

J Biol Chem 276:35087-35092 (2001)

PubMed id: 11445573

Crystal structure of a complex between Pseudomonas aeruginosa alkaline protease and its cognate inhibitor: inhibition by a zinc-NH2 coordinative bond.

T.Hege, R.E.Feltzer, R.D.Gray, U.Baumann.

ABSTRACT

Serralysins are a family of metalloproteases secreted by Gram-negative bacteria into the medium in the form of inactive zymogens. Usually, all serralysin secretors have on the same operon a gene coding for a periplasmic 10-kDa protein, which is an inhibitor of the secreted protease. The recent characterization of the inhibitor of the alkaline protease from Pseudomonas aeruginosa revealed a surprisingly low dissociation constant of 4 pm, contrary to earlier studies on homologous systems, where inhibition constants in the microm range were reported. To approach a more accurate understanding, the crystal structure of the complex between inhibitor and protease from P. aeruginosa was determined at 1.74 A resolution and refined to R(free) = 0.204. The structure reported here shows clearly that the N terminus of the inhibitor forms a coordinative bond to the catalytic Zn(2+) ion with a nitrogen-zinc distance of 2.17 A. We conclude that this interaction adds substantially to the complex stability and show also that similar interactions are found in other metzincin-inhibitor complexes.

Selected figure(s)

Figure 1.
Fig. 1. Structure of the APRin·APR complex. A, ribbon plot of the APR·APRin complex. The inhibitor is shown in red and the protease in cyan. The catalytic zinc is depicted as a magenta-colored ball. Green spheres symbolize calcium ions. This figure was prepared with the programs MOLSCRIPT (31) and Raster3d (32). B, surface representation showing the tight fit of the two solvent-accessible surfaces. Surfaces were calculated with MSMS and DINO (see web sites bioz.unibas.ch/~xray and scripps.edu/pub/olson-web/people/sanner/html/msms_man.html, respectively).

Figure 2.
Fig. 2. N terminus of APRin. A, stereo plot of the 2F[o] F[c] electron density map contoured at 1.5 . The inhibitor is shown in green and the protease in brown. B, the interactions of the N-terminal trunk of the inhibitor (green) with the protease (brown) and the zinc (yellow). The hydrogen bonds between Ser-2I and Glu-177P, Ser-2I and Ala-134P, and Leu-3I and Tyr-169P are shown as dotted black lines. Also shown are the hydrogen bonds between Ser-1I and Tyr-228P and Glu-177P. C, surface representation of APR together with the inhibitor model (carbons shown in green). The active site is closed by the flexible loop 188P-192P. This figure was prepared with MSMS and DINO.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2001, 276, 35087-35092) copyright 2001.

Figures were selected by the author.