The chaperone FdsC for Rhodobacter capsulatus formate dehydrogenase binds the bis-molybdopterin guanine dinucleotide cofactor.
Molybdoenzymes are complex enzymes in which the molybdenum cofactor (Moco) is deeply buried in the enzyme. Most molybdoenzymes contain a specific chaperone for the insertion of Moco. For the formate dehydrogenase FdsGBA from Rhodobacter capsulatus the two chaperones FdsC and FdsD were identified to be essential for enzyme activity, but are not a subunit of the mature enzyme. Here, we purified and characterized the FdsC protein after heterologous expression in Escherichia coli. We were able to copurify FdsC with the bound Moco derivate bis-molybdopterin guanine dinucleotide. This cofactor successfully was used as a source to reconstitute the activity of molybdoenzymes.
Institute of Biochemistry and Biology, Department of Molecular Enzymology, University of Potsdam, D-14476 Potsdam, Germany.
Nadine
N
Böhmer
Institute of Biochemistry and Biology, Department of Molecular Enzymology, University of Potsdam, D-14476 Potsdam, Germany.
Tobias
T
Hartmann
Institute of Biochemistry and Biology, Department of Molecular Enzymology, University of Potsdam, D-14476 Potsdam, Germany. Electronic address: sleim@uni-potsdam.de.
Silke
S
Leimkühler
D003067:Coenzymes / Q000378:metabolism*; D017124:Conserved Sequence; D005560:Formate Dehydrogenases / Q000378:metabolism*; D006150:Guanine Nucleotides / Q000378:metabolism*; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011622:Pterins / Q000378:metabolism*; D016354:Rhodobacter capsulatus / Q000201:enzymology*; D013379:Substrate Specificity
D003067:Coenzymes; D006150:Guanine Nucleotides; D018832:Molecular Chaperones; D011622:Pterins; C063761:molybdopterin guanine dinucleotide; D005560:Formate Dehydrogenases; D010089:Oxidoreductases, N-Demethylating; C023186:trimethylamine dehydrogenase
Chaperone; Formate dehydrogenase; Molybdenum cofactor; bis-MGD; l-cysteine desulfurase