| Biochemical and structural characterization of germicidin synthase: analysis of a type III polyketide synthase that employs acyl-ACP as a starter unit donor. | |
| | |
MedLine Citation:
|
PMID: 22480290 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Germicidin synthase (Gcs) from Streptomyces coelicolor is a type III polyketide synthase (PKS) with broad substrate flexibility for acyl groups linked through a thioester bond to either coenzyme A (CoA) or acyl carrier protein (ACP). Germicidin synthesis was reconstituted in vitro by coupling Gcs with fatty acid biosynthesis. Since Gcs has broad substrate flexibility, we directly compared the kinetic properties of Gcs with both acyl-ACP and acyl-CoA. The catalytic efficiency of Gcs for acyl-ACP was 10-fold higher than for acyl-CoA, suggesting a strong preference toward carrier protein starter unit transfer. The 2.9 Å germicidin synthase crystal structure revealed canonical type III PKS architecture along with an unusual helical bundle of unknown function that appears to extend the dimerization interface. A pair of arginine residues adjacent to the active site affect catalytic activity but not ACP binding. This investigation provides new and surprising information about the interactions between type III PKSs and ACPs that will facilitate the construction of engineered systems for production of novel polyketides. |
| | |
Authors:
|
Joseph A Chemler; Tonia J Buchholz; Todd W Geders; David L Akey; Christopher M Rath; George E Chlipala; Janet L Smith; David H Sherman |
Publication Detail:
|
Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2012-04-23 |
Journal Detail:
|
Title: Journal of the American Chemical Society Volume: 134 ISSN: 1520-5126 ISO Abbreviation: J. Am. Chem. Soc. Publication Date: 2012 May |
Date Detail:
|
Created Date: 2012-05-02 Completed Date: 2012-08-24 Revised Date: 2013-05-06 |
Medline Journal Info:
|
Nlm Unique ID: 7503056 Medline TA: J Am Chem Soc Country: United States |
Other Details:
|
Languages: eng Pagination: 7359-66 Citation Subset: IM |
Copyright Information:
|
© 2012 American Chemical Society |
Affiliation:
|
Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Acyl Carrier Protein
/
metabolism* Acyl Coenzyme A / metabolism Bacterial Proteins / chemistry, genetics, metabolism* Crystallography, X-Ray Models, Molecular Mutation Polyketide Synthases / chemistry, genetics, metabolism* Protein Binding Pyrones / metabolism* Signal Transduction Streptomyces coelicolor / chemistry, enzymology*, genetics Substrate Specificity |
| Grant Support | |
ID/Acronym/Agency:
|
DK42303/DK/NIDDK NIH HHS; GM076477/GM/NIGMS NIH HHS; GM095065/GM/NIGMS NIH HHS; R01 DK042303/DK/NIDDK NIH HHS; R01 GM076477/GM/NIGMS NIH HHS; R01 GM076477-07/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/Acyl Carrier Protein; 0/Acyl Coenzyme A; 0/Bacterial Proteins; 0/Pyrones; 151271-57-7/germicidin; 79956-01-7/Polyketide Synthases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Nucleophilic Substitution: A Charge Density Perspective.
Next Document: RP-HPLC method for the simultaneous quantitation of boeravinone E and boeravinone B in Boerhaavia di...