Expression of an active adenylate-forming domain of peptide synthetases corresponding to acyl-CoA-synthetases.
Peptide synthetases and acyl-CoA-synthetases form acyl adenylates which are transferred to CoA or enzyme-bound pantetheine. To verify the existence of an adenylate domain in peptide synthetases, a 60.8 kDa fragment of tyrocidine 1-synthetase was constructed by a 1,629 bp deletion, expressed in Escherichia coli, and characterized. The truncated multienzyme activated phenylalanine and substrate analogues with comparable kinetics as the over-expressed synthetase, as judged by ATP-[32P]PP(i) exchange reaction. Thus the N-terminal domain resembling an acyl-CoA-synthetase is an autonomous structural element. This N-terminal domain is followed by a cofactor binding domain, resembling acyl carrier proteins involved in polyketide formation.
Dieckmann
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R
Institut für Biochemie und Molekulare Biologie, Technische Universität Berlin, Germany.
Lee
Y O
YO
van Liempt
H
H
von Döhren
H
H
Kleinkauf
H
H
D000249:Adenosine Monophosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D001665:Binding Sites; D003066:Coenzyme A Ligases / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli; D008969:Molecular Sequence Data; D010453:Peptide Synthases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010649:Phenylalanine / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D011994:Recombinant Proteins; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid
D000596:Amino Acids; D010710:Phosphates; D011994:Recombinant Proteins; D000249:Adenosine Monophosphate; D010649:Phenylalanine; D000255:Adenosine Triphosphate; D003066:Coenzyme A Ligases; D010453:Peptide Synthases; C035180:tyrocidine synthetase