Dieckmann R, Lee Y O, van Liempt H, von Döhren H, Kleinkauf H
Institut für Biochemie und Molekulare Biologie, Technische Universität Berlin, Germany.
FEBS Lett. 1995 Jan 3;357(2):212-6. doi: 10.1016/0014-5793(94)01342-x.
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.
肽合成酶和酰基辅酶A合成酶形成酰基腺苷酸，其被转移至辅酶A或与酶结合的泛酰巯基乙胺。为验证肽合成酶中腺苷酸结构域的存在，通过1629 bp的缺失构建了酪蛋白1合成酶的60.8 kDa片段，在大肠杆菌中表达并进行表征。通过ATP-[32P]PP(i)交换反应判断，截短的多酶激活苯丙氨酸和底物类似物的动力学与过表达的合成酶相当。因此，类似于酰基辅酶A合成酶的N端结构域是一个自主的结构元件。该N端结构域之后是一个辅因子结合结构域，类似于参与聚酮形成的酰基载体蛋白。
