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Acylation of streptomyces type II polyketide synthase acyl carrier proteins

Acylation of streptomyces type II polyketide synthase acyl carrier proteins
Acylation of streptomyces type II polyketide synthase acyl carrier proteins
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.
0014-5793
132-138
Crosby, John
38dff705-47dc-4406-be2b-cce3599a776b
Byrom, Kate J.
6dc62fc3-4c01-4552-81c1-ac3cb1f8276b
Hitchman, Timothy S.
3040aef9-0663-4872-9ae2-f5af27ab87a6
Cox, Russell J.
ddcf3a6a-955e-4f54-9809-f497b189468b
Crump, Matthew P.
ed31b5fd-23f6-434c-a38c-da3cb7b27402
Findlow, I. Stuart C.
e32d6c84-a6ec-42d8-bb05-a8411b48c856
Bibb, Maureen J.
73563bae-c361-4711-a7b2-dc9b1c34ccbd
Simpson, Thomas J.
17c403f6-38f4-4c81-9267-319f2d441183
Crosby, John
38dff705-47dc-4406-be2b-cce3599a776b
Byrom, Kate J.
6dc62fc3-4c01-4552-81c1-ac3cb1f8276b
Hitchman, Timothy S.
3040aef9-0663-4872-9ae2-f5af27ab87a6
Cox, Russell J.
ddcf3a6a-955e-4f54-9809-f497b189468b
Crump, Matthew P.
ed31b5fd-23f6-434c-a38c-da3cb7b27402
Findlow, I. Stuart C.
e32d6c84-a6ec-42d8-bb05-a8411b48c856
Bibb, Maureen J.
73563bae-c361-4711-a7b2-dc9b1c34ccbd
Simpson, Thomas J.
17c403f6-38f4-4c81-9267-319f2d441183

Crosby, John, Byrom, Kate J., Hitchman, Timothy S., Cox, Russell J., Crump, Matthew P., Findlow, I. Stuart C., Bibb, Maureen J. and Simpson, Thomas J. (1998) Acylation of streptomyces type II polyketide synthase acyl carrier proteins. FEBS Letters, 433 (1-2), 132-138. (doi:10.1016/S0014-5793(98)00840-0).

Record type: Article

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.

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Published date: 14 August 1998

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Local EPrints ID: 156641
URI: http://eprints.soton.ac.uk/id/eprint/156641
ISSN: 0014-5793
PURE UUID: 180fbf8a-0e54-405c-9a3d-5b5ce84c3296

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Date deposited: 01 Jun 2010 12:21
Last modified: 14 Mar 2024 01:44

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Contributors

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

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