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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
46
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pubmed:dateCreated |
2005-11-15
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pubmed:abstractText |
During polyketide biosynthesis, malonyl groups are transferred to the acyl carrier protein (ACP) component of the polyketide synthase (PKS), and it has been shown that a number of type II polyketide ACPs undergo rapid self-acylation from malonyl-CoA in the absence of a malonyl-CoA:holo-acyl carrier protein transacylase (MCAT). More recently, however, the observation of self-malonylation has been ascribed to contamination with Escherichia coli MCAT (FabD) rather than an intrinsic property of the ACP. The wild-type apo-ACP from the actinorhodin (act) PKS of Streptomyces coelicolor (synthetic apo-ACP) has therefore been synthesized using solid-state peptide methods and refolded using the GroEL/ES chaperone system from E. coli. Correct folding of the act ACP has been confirmed by circular dichroism (CD) and 1H NMR. Synthetic apo-ACP was phosphopantetheinylated to 100% by S. coelicolor holo-acyl carrier protein synthase (ACPS), and the resultant holo-ACP underwent self-malonylation in the presence of malonyl-CoA. No malonylation of negative controls was observed, confirming that the use of ACPS and GroEL/ES did not introduce contamination with E. coli MCAT. This result proves unequivocally that self-malonylation is an inherent activity of this PKS ACP in vitro.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0006-2960
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
22
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pubmed:volume |
44
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
15414-21
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:16285746-Acyl Carrier Protein,
pubmed-meshheading:16285746-Acyl-Carrier Protein S-Malonyltransferase,
pubmed-meshheading:16285746-Apoproteins,
pubmed-meshheading:16285746-Chromatography, High Pressure Liquid,
pubmed-meshheading:16285746-Circular Dichroism,
pubmed-meshheading:16285746-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:16285746-Malonates,
pubmed-meshheading:16285746-Nuclear Magnetic Resonance, Biomolecular,
pubmed-meshheading:16285746-Polyketide Synthases,
pubmed-meshheading:16285746-Protein Folding,
pubmed-meshheading:16285746-Spectrometry, Mass, Electrospray Ionization,
pubmed-meshheading:16285746-Streptomyces coelicolor
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pubmed:year |
2005
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pubmed:articleTitle |
Self-malonylation is an intrinsic property of a chemically synthesized type II polyketide synthase acyl carrier protein.
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pubmed:affiliation |
School of Chemistry, University of Bristol, Cantock's Close, Bristol BS8 1TS, UK.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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