Premium
Crystal structure of the hydroxylase component of methane monooxygenase from Methylosinus trichosporium OB3b
Author(s) -
Elango Nates An,
Radhakrishnan Ramaswamy,
Froland Wayne A.,
Wallar Bradley J.,
Earhart Cathleen A.,
Lipscomb John D.,
Ohlendorf Douglas H.
Publication year - 1997
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560060305
Subject(s) - methane monooxygenase , chemistry , carboxylate , stereochemistry , crystal structure , oxidoreductase , active site , methanotroph , crystallography , enzyme , methane , anaerobic oxidation of methane , biochemistry , organic chemistry
Abstract Methane monooxygenase (MMO), found in aerobic methanotrophic bacteria, catalyzes the 02‐dependent conversion of methane to methanol. The soluble form of the enzyme (sMMO) consists of three components: a reductase, a regulatory “B” component (MMOB), and a hydroxylase component (MMOH), which contains a hydroxo‐bridged dinuclear iron cluster. Two genera of methanotrophs, termed Type X and Type II, which differ markedly in cellular and metabolic characteristics, are known to produce the sMMO. The structure of MMOH from the Type X methanotroph Methylo‐coccus capsulatus Bath (MMO Bath) has been reported recently. Two different structures were found for the essential diiron cluster, depending upon the temperature at which the diffraction data were collected. In order to extend the structural studies to the Type II methanotrophs and to determine whether one of the two known MMOH structures is generally applicable to the MMOH family, we have determined the crystal structure of the MMOH from Type II Methylosinus trichosporium OB3b (MMO OB3b) in two crystal forms to 2.0 A and 2.7 A resolution, respectively, both determined at 18 °C. The crystal forms differ in that MMOB was present during crystallization of the second form. Both crystal forms, however, yielded very similar results for the structure of the MMOH. Most of the major structural features of the MMOH Bath were also maintained with high fidelity. The two irons of the active site cluster of MMOH OB3b are bridged by two OH (or one OH and one H20), as well as both carboxylate oxygens of Glu a 144. This bis‐yu.‐hydroxo‐bridged “diamond core” structure, with a short Fe‐Fe distance of 2.99 A, is unique for the resting state of proteins containing analogous diiron clusters, and is very similar to the structure reported for the cluster from flash frozen (‐160°C) crystals of MMOH Bath, suggesting a common active site structure for the soluble MMOHs. The high‐resolution structure of MMOH OB3b indicates 26 consecutive amino acid sequence differences in the /3 chain when compared to the previously reported sequence inferred from the cloned gene. Fifteen additional sequence differences distributed randomly over the three chains were also observed, including Da209E, a ligand of one of the irons.