Acylation of Streptomyces type II polyketide synthase acyl carrier proteins

John Crosby; Kate J. Byrom; Timothy S. Hitchman; Russell J. Cox; Matthew P. Crump; I. Stuart C. Findlow; Maureen J. Bibb; Thomas J. Simpson

doi:10.1016/S0014-5793(98)00840-0

Acylation of Streptomyces type II polyketide synthase acyl carrier proteins

John Crosby
, Kate J. Byrom
, Timothy S. Hitchman
, Russell J. Cox
, Matthew P. Crump
, I. Stuart C. Findlow
, Maureen J. Bibb
, Thomas J. Simpson^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Research › peer review

Abstract

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. Copyright (C) 1998 Federation of European Biochemical Societies.

Original language	English
Pages (from-to)	132-138
Number of pages	7
Journal	FEBS letters
Volume	433
Issue number	1-2
DOIs	https://doi.org/10.1016/S0014-5793(98)00840-0
Publication status	Published - 14 Aug 1998
Externally published	Yes

Keywords

Acyl carrier protein
Acylation
Fatty acid synthase
Polyketide synthase

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/S0014-5793(98)00840-0

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title = "Acylation of Streptomyces type II polyketide synthase acyl carrier proteins",

abstract = "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. Copyright (C) 1998 Federation of European Biochemical Societies.",

keywords = "Acyl carrier protein, Acylation, Fatty acid synthase, Polyketide synthase",

author = "John Crosby and Byrom, \{Kate J.\} and Hitchman, \{Timothy S.\} and Cox, \{Russell J.\} and Crump, \{Matthew P.\} and Findlow, \{I. Stuart C.\} and Bibb, \{Maureen J.\} and Simpson, \{Thomas J.\}",

note = "Funding information: This work was supported by the Engineering and Physical Sciences Research Council (EPSRC), the Biotechnology and Biological Sciences Research Council (BBSRC) and the John Innes Foundation. Studentships from the EPSRC (I.S.C.F., T.S.H. and M.P.C.) and School of Chemistry (K.J.B.) are gratefully acknowledged. R.J.C. gratefully acknowledges the receipt of a Royal Society equipment grant.",

year = "1998",

month = aug,

day = "14",

doi = "10.1016/S0014-5793(98)00840-0",

language = "English",

volume = "433",

pages = "132--138",

journal = "FEBS letters",

issn = "0014-5793",

publisher = "John Wiley and Sons Inc.",

number = "1-2",

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T1 - Acylation of Streptomyces type II polyketide synthase acyl carrier proteins

AU - Crosby, John

AU - Byrom, Kate J.

AU - Hitchman, Timothy S.

AU - Cox, Russell J.

AU - Crump, Matthew P.

AU - Findlow, I. Stuart C.

AU - Bibb, Maureen J.

AU - Simpson, Thomas J.

N1 - Funding information: This work was supported by the Engineering and Physical Sciences Research Council (EPSRC), the Biotechnology and Biological Sciences Research Council (BBSRC) and the John Innes Foundation. Studentships from the EPSRC (I.S.C.F., T.S.H. and M.P.C.) and School of Chemistry (K.J.B.) are gratefully acknowledged. R.J.C. gratefully acknowledges the receipt of a Royal Society equipment grant.

PY - 1998/8/14

Y1 - 1998/8/14

N2 - 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. Copyright (C) 1998 Federation of European Biochemical Societies.

AB - 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. Copyright (C) 1998 Federation of European Biochemical Societies.

KW - Acyl carrier protein

KW - Acylation

KW - Fatty acid synthase

KW - Polyketide synthase

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U2 - 10.1016/S0014-5793(98)00840-0

DO - 10.1016/S0014-5793(98)00840-0

M3 - Article

C2 - 9738947

AN - SCOPUS:0031872017

SN - 0014-5793

VL - 433

SP - 132

EP - 138

JO - FEBS letters

JF - FEBS letters

IS - 1-2

ER -

Acylation of Streptomyces type II polyketide synthase acyl carrier proteins

Abstract

Keywords

ASJC Scopus subject areas

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