PDBsum entry 1oyv

Hydrolase

PDB id: 1oyv

Contents

Protein chains

274 a.a. *

104 a.a. *

Metals

_CA ×2

Waters ×128

* Residue conservation analysis

PDB id:

1oyv

Name:

Hydrolase

Title:

Crystal structure of tomato inhibitor-ii in a ternary complex with subtilisin carlsberg

Structure:

Subtilisin carlsberg. Chain: a, b. Wound-induced proteinase inhibitor-ii. Chain: i

Source:

Bacillus licheniformis. Organism_taxid: 1402. Strain: nagarse. Solanum lycopersicum. Organism_taxid: 4081. Strain: bonny best. Other_details: leaves of tomato plant

Biol. unit:

Trimer (from PQS)

Resolution:

2.50Å R-factor: 0.214 R-free: 0.270

Authors:

I.H.Barrette-Ng,K.K.Ng,M.M.Cherney,G.Pearce,C.A.Ryan,M.N.James

Key ref:

I.H.Barrette-Ng et al. (2003). Structural basis of inhibition revealed by a 1:2 complex of the two-headed tomato inhibitor-II and subtilisin Carlsberg. J Biol Chem, 278, 24062-24071. PubMed id: 12684499 DOI: 10.1074/jbc.M302020200

Date:

07-Apr-03 Release date: 15-Jul-03

Protein chains

P00780  (SUBC_BACLI) -  Subtilisin Carlsberg from Bacillus licheniformis

Seq: 379 a.a.

Struc: 274 a.a.*

Protein chain

P05119  (IP21_SOLLC) -  Wound-induced proteinase inhibitor 2 from Solanum lycopersicum

Seq: 148 a.a.

Struc: 104 a.a.

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

Enzyme reactions

• Enzyme class: Chains A, B: E.C.3.4.21.62 - subtilisin.
• Reaction: Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyzes peptide amides.

DOI no: 10.1074/jbc.M302020200

J Biol Chem 278:24062-24071 (2003)

PubMed id: 12684499

Structural basis of inhibition revealed by a 1:2 complex of the two-headed tomato inhibitor-II and subtilisin Carlsberg.

I.H.Barrette-Ng, K.K.Ng, M.M.Cherney, G.Pearce, C.A.Ryan, M.N.James.

ABSTRACT

Multidomain proteinase inhibitors play critical roles in the defense of plants against predation by a wide range of pests. Despite a wealth of structural information on proteinase-single domain inhibitor interactions, the structural basis of inhibition by multidomain proteinase inhibitors remains poorly understood. Here we report the 2.5-A resolution crystal structure of the two-headed tomato inhibitor-II (TI-II) in complex with two molecules of subtilisin Carlsberg; it reveals how a multidomain inhibitor from the Potato II family of proteinase inhibitors can bind to and simultaneously inhibit two enzyme molecules within a single ternary complex. The N terminus of TI-II initiates the folding of Domain I (Lys-1 to Cys-15 and Pro-84 to Met-123) and then completes Domain II (Ile-26 to Pro-74) before coming back to complete the rest of Domain I (Pro-84 to Met-123). The two domains of TI-II adopt a similar fold and are arranged in an extended configuration that presents two reactive site loops at the opposite ends of the inhibitor molecule. Each subtilisin molecule interacts with a reactive site loop of TI-II through the standard, canonical binding mode. Remarkably, a significant distortion of the active site of subtilisin is induced by the presence of phenylalanine in the P1 position of reactive site loop II of TI-II. The structure of the TI-II.(subtilisin)2 complex provides a molecular framework for understanding how multiple inhibitory domains in a single Potato II type proteinase inhibitor molecule from the Potato II family act to inhibit proteolytic enzymes.

Selected figure(s)

Figure 4.
FIG. 4. Conformations of the reactive site loops. A, stereoscopic view of a superposition of the reactive site loops of both domains. Conformation of the reactive site loops of Domain I (B) and Domain II (C) bound to subtilisin are shown. The solvent-accessible surface of the subtilisin molecules are drawn with negatively charged residues colored red, positively charged residues colored blue, and hydrophobic residues colored magenta.

Figure 5.
FIG. 5. Interactions between subtilisin and TI-II. Shown is a stylized representation of the interactions between subtilisin and the reactive site loops of Domain I (A) and Domain II (B). Residues of subtilisin making van der Waals interactions with the reactive site loops of TI-II are shown as blue circles and, in some instances, as parallel lines. Also shown is a stereoscopic view of the interactions between subtilisin and the reactive site loops of Domain I (C) and Domain II (D). Inhibitor is green, and subtilisin is black.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2003, 278, 24062-24071) copyright 2003.

Figures were selected by an automated process.