Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 685-692 |
| Seitenumfang | 8 |
| Fachzeitschrift | Nature chemical biology |
| Jahrgang | 9 |
| Ausgabenummer | 11 |
| Publikationsstatus | Veröffentlicht - Nov. 2013 |
| Extern publiziert | Ja |
Abstract
Type I polyketide synthases often use programmed β-branching, via enzymes of a 'hydroxymethylglutaryl-CoA synthase (HCS) cassette', to incorporate various side chains at the second carbon from the terminal carboxylic acid of growing polyketide backbones. We identified a strong sequence motif in acyl carrier proteins (ACPs) where β-branching is known to occur. Substituting ACPs confirmed a correlation of ACP type with β-branching specificity. Although these ACPs often occur in tandem, NMR analysis of tandem β-branching ACPs indicated no ACP-ACP synergistic effects and revealed that the conserved sequence motif forms an internal core rather than an exposed patch. Modeling and mutagenesis identified ACP helix III as a probable anchor point of the ACP-HCS complex whose position is determined by the core. Mutating the core affects ACP functionality, whereas ACP-HCS interface substitutions modulate system specificity. Our method for predicting β-carbon branching expands the potential for engineering new polyketides and lays a basis for determining specificity rules.
ASJC Scopus Sachgebiete
- Biochemie, Genetik und Molekularbiologie (insg.)
- Molekularbiologie
- Biochemie, Genetik und Molekularbiologie (insg.)
- Zellbiologie
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in: Nature chemical biology, Jahrgang 9, Nr. 11, 11.2013, S. 685-692.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - A conserved motif flags acyl carrier proteins for β-branching in polyketide synthesis
AU - Haines, Anthony S.
AU - Dong, Xu
AU - Song, Zhongshu
AU - Farmer, Rohit
AU - Williams, Christopher
AU - Hothersall, Joanne
AU - Płoskoń, Eliza
AU - Wattana-Amorn, Pakorn
AU - Stephens, Elton R.
AU - Yamada, Erika
AU - Gurney, Rachel
AU - Takebayashi, Yuiko
AU - Masschelein, Joleen
AU - Cox, Russell J.
AU - Lavigne, Rob
AU - Willis, Christine L.
AU - Simpson, Thomas J.
AU - Crosby, John
AU - Winn, Peter J.
AU - Thomas, Christopher M.
AU - Crump, Matthew P.
N1 - Funding information: This work was supported by the Biotechnology and Biological Sciences Research Council and the Engineering and Physical Sciences Research Council (BB/F014570/1 to P.W-a. and X.D.; BB/I014373/1 to A.S.H., R.G. and J.H.; EP/F066104/1, BB/I003355/1 and BB/I014039/1 to Z.S. and for LC/MS equipment). E.P. was supported by a European Union studentship grant (FP6-mobility 504501). R.F. and Y.T. were supported by scholarships from the Darwin Trust of Edinburgh. J.M. is supported by a PhD scholarship from the FWO Vlaanderen. P.J.W. and R.F. thank A. Bonvin for technical assistance with HADDOCK.
PY - 2013/11
Y1 - 2013/11
N2 - Type I polyketide synthases often use programmed β-branching, via enzymes of a 'hydroxymethylglutaryl-CoA synthase (HCS) cassette', to incorporate various side chains at the second carbon from the terminal carboxylic acid of growing polyketide backbones. We identified a strong sequence motif in acyl carrier proteins (ACPs) where β-branching is known to occur. Substituting ACPs confirmed a correlation of ACP type with β-branching specificity. Although these ACPs often occur in tandem, NMR analysis of tandem β-branching ACPs indicated no ACP-ACP synergistic effects and revealed that the conserved sequence motif forms an internal core rather than an exposed patch. Modeling and mutagenesis identified ACP helix III as a probable anchor point of the ACP-HCS complex whose position is determined by the core. Mutating the core affects ACP functionality, whereas ACP-HCS interface substitutions modulate system specificity. Our method for predicting β-carbon branching expands the potential for engineering new polyketides and lays a basis for determining specificity rules.
AB - Type I polyketide synthases often use programmed β-branching, via enzymes of a 'hydroxymethylglutaryl-CoA synthase (HCS) cassette', to incorporate various side chains at the second carbon from the terminal carboxylic acid of growing polyketide backbones. We identified a strong sequence motif in acyl carrier proteins (ACPs) where β-branching is known to occur. Substituting ACPs confirmed a correlation of ACP type with β-branching specificity. Although these ACPs often occur in tandem, NMR analysis of tandem β-branching ACPs indicated no ACP-ACP synergistic effects and revealed that the conserved sequence motif forms an internal core rather than an exposed patch. Modeling and mutagenesis identified ACP helix III as a probable anchor point of the ACP-HCS complex whose position is determined by the core. Mutating the core affects ACP functionality, whereas ACP-HCS interface substitutions modulate system specificity. Our method for predicting β-carbon branching expands the potential for engineering new polyketides and lays a basis for determining specificity rules.
UR - http://www.scopus.com/inward/record.url?scp=84886596825&partnerID=8YFLogxK
U2 - 10.1038/nchembio.1342
DO - 10.1038/nchembio.1342
M3 - Article
AN - SCOPUS:84886596825
VL - 9
SP - 685
EP - 692
JO - Nature chemical biology
JF - Nature chemical biology
SN - 1552-4450
IS - 11
ER -