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Carbamoylphosphate serves as the source of CN − , but not of the intrinsic CO in the active site of the regulatory [NiFe]‐hydrogenase from Ralstonia eutropha
Author(s) -
Lenz Oliver,
Zebger Ingo,
Hamann Josta,
Hildebrandt Peter,
Friedrich Bärbel
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.06.027
Subject(s) - ralstonia , hydrogenase , chemistry , cyanide , ligand (biochemistry) , active site , covalent bond , cupriavidus necator , formate , stereochemistry , catalysis , biochemistry , combinatorial chemistry , enzyme , organic chemistry , bacteria , biology , receptor , polyhydroxyalkanoates , genetics
Within the catalytic centre of [NiFe]‐hydrogenases one carbonyl and two cyanide ligands are covalently attached to the iron. To identify the metabolic origins of these ligands, the regulatory [NiFe] hydrogenase in conjunction with the indigenous Hyp maturation proteins of Ralstonia eutropha H16 were heterologously overproduced in E. coli grown in the presence of l ‐[ureido‐ 13 C] citrulline and NaH 13 CO 3 . Infrared spectroscopy of purified hydrogenase provided direct evidence that only the cyanide ligands, but not the CO ligand, originate from CO 2 and carbamoylphosphate. Incorporation of label from 13 CO exclusively into the carbonyl ligand indicates that free CO is a possible precursor in carbonyl ligand biosynthesis.