6-S-cysteinyl flavin mononucleotide-containing histamine dehydrogenase from Nocardioides simplex: molecular cloning, sequencing, overexpression, and characterization of redox centers of enzyme.
Histamine dehydrogenase from Nocardioides simplex is a homodimeric enzyme and catalyzes oxidative deamination of histamine. The gene encoding this enzyme has been sequenced and cloned by polymerase chain reactions and overexpressed in Escherichia coli. The sequence of the complete open reading frame, 2073 bp coding for a protein of 690 amino acids, was determined on both strands. The amino acid sequence of histamine dehydrogenase is closely related to those of trimethylamine dehydrogenase and dimethylamine dehydrogenase containing an unusual covalently bound flavin mononucleotide, 6-S-cysteinyl-flavin mononucleotide, and one 4Fe-4S cluster as redox active cofactors in each subunit of the homodimer. The presence of the identical redox cofactors in histamine dehydrogenase has been confirmed by sequence alignment analysis, mass spectral analysis, UV-vis and EPR spectroscopy, and chemical analysis of iron and acid-labile sulfur. These results suggest that the structure of histamine dehydrogenase in the vicinity of the two redox centers is almost identical to that of trimethylamine dehydrogenase as a whole. The structure modeling study, however, demonstrated that a putative substrate-binding cavity in histamine dehydrogenase is quite distinct from that of trimethylamine dehydrogenase.
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
Nobutaka
N
Fujieda
Atsuko
A
Satoh
Noriaki
N
Tsuse
Kenji
K
Kano
Tokuji
T
Ikeda
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry; D001665:Binding Sites; D020134:Catalytic Domain; D003001:Cloning, Molecular; D005486:Flavin Mononucleotide / Q000737:chemistry; D006094:Gram-Positive Bacteria / Q000201:enzymology*; D007506:Iron-Sulfur Proteins / Q000737:chemistry; D008958:Models, Molecular; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D010089:Oxidoreductases, N-Demethylating / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D013057:Spectrum Analysis
D001426:Bacterial Proteins; D007506:Iron-Sulfur Proteins; D005486:Flavin Mononucleotide; D000587:Oxidoreductases Acting on CH-NH Group Donors; D010089:Oxidoreductases, N-Demethylating; C023186:trimethylamine dehydrogenase; C020064:dimethylamine dehydrogenase; C490533:histamine dehydrogenase, Nocardioides simplex