PDBsum entry 1vsb

Serine protease

PDB id: 1vsb

Contents

Protein chain

274 a.a. *

Waters ×84

* Residue conservation analysis

PDB id:

1vsb

Name:

Serine protease

Title:

Subtilisin carlsberg l-para-chlorophenyl-1-acetamido boronic acid inhibitor complex

Structure:

Subtilisin carlsberg, type viii. Chain: a. Synonym: subtilopeptidase a. Ec: 3.4.21.62

Source:

Bacillus licheniformis. Organism_taxid: 1402. Other_details: purchased from sigma

Resolution:

2.10Å R-factor: 0.171 R-free: 0.242

Authors:

V.S.Stoll,B.T.Eger,R.C.Hynes,V.Martichonok,J.B.Jones,E.F.Pai

Key ref:

V.S.Stoll et al. (1998). Differences in binding modes of enantiomers of 1-acetamido boronic acid based protease inhibitors: crystal structures of gamma-chymotrypsin and subtilisin Carlsberg complexes. Biochemistry, 37, 451-462. PubMed id: 9425066 DOI: 10.1021/bi971166o

Date:

17-Sep-97 Release date: 18-Mar-98

Protein chain

P00780  (SUBC_BACLI) -  Subtilisin Carlsberg from Bacillus licheniformis

Seq: 379 a.a.

Struc: 274 a.a.*

* PDB and UniProt seqs differ at 4 residue positions (black crosses)

Enzyme reactions

• Enzyme class: 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: 10.1021/bi971166o

Biochemistry 37:451-462 (1998)

PubMed id: 9425066

Differences in binding modes of enantiomers of 1-acetamido boronic acid based protease inhibitors: crystal structures of gamma-chymotrypsin and subtilisin Carlsberg complexes.

V.S.Stoll, B.T.Eger, R.C.Hynes, V.Martichonok, J.B.Jones, E.F.Pai.

ABSTRACT

In order to probe the structural basis of stereoselectivity in the serine protease family, a series of enantiomeric boronic acids RCH2CH(NHCOCH3)B(OH)2 has been synthesized and kinetically characterized as transition-state analog inhibitors using alpha-chymotrypsin and subtilisin Carlsberg as model systems. When the R-substituent in this series was changed from a p-chlorophenyl to a 1-naphthyl group, alpha-chymotrypsin, but not subtilisin, reversed its usual preference for l-enantiomers and bound more tightly to the D-enantiomer [Martichonok, V., & Jones, J. B. (1996) J. Am. Chem. Soc. 118, 950-958]. The structural factors responsible for the differences in stereoselectivity between the two enzymes have been explored by X-ray crystallographic examination of subtilisin Carlsberg and gamma-chymotrypsin complexes of the L- and D-enantiomers of p-chlorophenyl and 1-naphthyl boronic acid derivatives. In both enzymes, the L-isomers of the inhibitors, which are more closely related to the natural L-amino acid substrates, form tetrahedral adducts, covalently linking the central boron atom and Ogamma of the catalytic serine. The d-isomers, however, differ in the way they interact with subtilisin or gamma-chymotrypsin. With subtilisin, both the D-p-chlorophenyl and D-1-naphthyl inhibitor complexes form covalent Ser Ogamma-to-boron bonds, but with gamma-chymotrypsin, the same inhibitors lead to novel tetrahedral adducts covalently linking both Ser195 Ogamma and His57 Nepsilon2 covalently via the boron atom.