Open Accessp ‐Hydroxybenzoate Hydroxylase from Pseudomonas fluorescens
Author(s) -
WEIJER Wicher J.,
HOFSTEENGE Jan,
BEINTEMA Jaap J.,
WIERENGA Rik K.,
DRENTH Jan
Publication year - 1983
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1983.tb07435.x
Subject(s) - side chain , protein tertiary structure , stereochemistry , substrate (aquarium) , chemistry , pseudomonas fluorescens , hydroxybenzoate , protein subunit , protein primary structure , amino acid , peptide sequence , binding site , active site , enzyme , biochemistry , biology , organic chemistry , bacteria , ecology , genetics , gene , polymer
The complete primary and tertiary structure of p ‐hydroxybenzoate hydroxylase is now known. The amino acid sequences of the two largest CNBr peptides have been fitted to the electron‐density map at 0.25‐nm resolution. The parts of the polypeptide chain contributing the residues to the FAD‐binding site and the residues of the substrate‐binding site have been identified. The active site is located in a large hydrophobic area enclosed by all domains of the enzyme structure. Here the substrate, p ‐hydroxybenzoate, is bound near, but not in direct contact with, the isoalloxazine ring system of FAD. Many side chains from the C‐terminal part of the polypeptide chain are involved in subunit‐subunit interactions. In the center of one of the largely hydrophobic contact areas between the subunits, a cluster of six aromatic amino acids was found.