Open AccessExpression, purification, crystallization and preliminary X‐ray studies of histamine dehydrogenase from Nocardioides simplex
Author(s) -
Reed Timothy M.,
Hirakawa Hidehiko,
Mure Minae,
Scott Emily E.,
Limburg Julian
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108023336
Subject(s) - crystallization , chemistry , simplex , histamine , biochemistry , biology , organic chemistry , pharmacology , geometry , mathematics
Histamine dehydrogenase (HADH) from Nocardioides simplex catalyzes the oxidative deamination of histamine to produce imidazole acetaldehyde and an ammonium ion. HADH is functionally related to trimethylamine dehydrogenase (TMADH), but HADH has strict substrate specificity towards histamine. HADH is a homodimer, with each 76 kDa subunit containing two redox cofactors: a [4Fe–4S] cluster and an unusual covalently bound flavin mononucleotide, 6‐ S ‐cysteinyl‐FMN. In order to understand the substrate specificity of HADH, it was sought to determine its structure by X‐ray crystallography. This enzyme has been expressed recombinantly in Escherichia coli and successfully crystallized in two forms. Diffraction data were collected to 2.7 Å resolution at the SSRL synchrotron with 99.7% completeness. The crystals belonged to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 101.14, b = 107.03, c = 153.35 Å.