Acylation of Streptomyces type II polyketide synthase acyl carrier proteins

FEBS Letters

Cox, Russell J²  Crosby, John²  Simpson, Thomas J²  Bibb, Maureen J¹  Byrom, Kate J²  Hitchman, Timothy S²  Crump, Matthew P²  Findlow, I.Stuart C² 

[1] John Innes Centre, Colney Lane, Norwich NR4 7UH, UK;School of Chemistry, University of Bristol, Cantock's Close, Bristol BS8 1TS, UK

关键词: Acyl carrier protein;    Acylation;    Polyketide synthase;    Fatty acid synthase;    ACP;    acyl carrier protein;    act;    actinorhodin;    ESMS;    electrospray mass spectrometry;    FAS;    fatty acid synthase;    gris;    griseusin;    NPDS;    p-nitrophenyl disulphide;    NPS;    nitrophenyl sulphide;    PKS;    polyketide synthase;    otc;    oxytetracycline;   

DOI  :  10.1016/S0014-5793(98)00840-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Acyl derivatives of type II PKS ACPs are required for in vitro studies of polyketide biosynthesis. The presence of an exposed cysteine residue prevented specific chemical acylation of the phosphopantetheine thiol of the actinorhodin PKS holo ACP. Acylation studies were further complicated by intramolecular disulphide formation between cysteine 17 and the phosphopantetheine. The presence of this intramolecular disulphide was confirmed by tryptic digestion of the ACP followed by ESMS analysis of the fragments. An act Cys17Ser ACP was engineered by site-directed mutagenesis. S-Acyl adducts of act C17S, oxytetracycline and griseusin holo ACPs were rapidly formed by reaction with hexanoyl, 5-ketohexanoyl and protected acetoacetyl imidazolides. Comparisons with type II FAS ACPs were made.

【 授权许可】

Unknown