PDBsum entry 2vgc

Serine protease

PDB id: 2vgc

Contents

Protein chains

131 a.a. *

95 a.a. *

Ligands

CYS-GLY-VAL-PRO-ALA-ILE-GLN-PRO-VAL-LEU

SO4 ×2

V35

Waters ×95

* Residue conservation analysis

References listed in PDB file

Key reference

• Title: Differences in binding modes of enantiomers of 1-Acetamido boronic acid based protease inhibitors: crystal structures of gamma-Chymotrypsin and subtilisin carlsberg complexes.
• Authors: V.S.Stoll, B.T.Eger, R.C.Hynes, V.Martichonok, J.B.Jones, E.F.Pai.
• Ref.: Biochemistry, 1998, 37, 451-462. [DOI no: 10.1021/bi971166o]
• PubMed id: 9425066

Note In the PDB file this reference is annotated as "TO BE PUBLISHED". The citation details given above were identified by an automated search of PubMed on title and author names, giving a percentage match of 96%.

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.

Secondary reference #1

• Title: Probing the specificity of the serine proteases subtilisin carlsberg and a-Chymotrypsin with enantiomeric 1-Acetamido boronic acids. An unexpected reversal of the normal
• Authors: V.Martichonok, J.B.Jones.
• Ref.: j am chem soc, 1996, 118, 95.

Secondary reference #2

• Title: Probing the specificity of the s1 binding site of subtilisin carlsberg with boronic acids.
• Authors: P.Seufer-Wasserthal, V.Martichonok, T.H.Keller, B.Chin, R.Martin, J.B.Jones.
• Ref.: Bioorg Med Chem Lett, 1994, 2, 35-48.
• PubMed id: 7922119

Secondary reference #3

• Title: Gamma-Chymotrypsin is a complex of alpha-Chymotrypsin with its own autolysis products.
• Authors: M.Harel, C.T.Su, F.Frolow, I.Silman, J.L.Sussman.
• Ref.: Biochemistry, 1991, 30, 5217-5225. [DOI no: 10.1021/bi00235a015]
• PubMed id: 2036388

Secondary reference #4

• Title: Structure and activity of two photoreversible cinnamates bound to chymotrypsin.
• Authors: B.L.Stoddard, J.Bruhnke, N.Porter, D.Ringe, G.A.Petsko.
• Ref.: Biochemistry, 1990, 29, 4871-4879. [DOI no: 10.1021/bi00472a017]
• PubMed id: 2364065