Molecular basis of Celmer's rules: Stereochemistry of catalysis by isolated ketoreductase domains from modular polyketide synthases

Siskos, Alexandros P., Baerga-Ortiz, Abel, Bali, Shilpa, Stein, Viktor, Mamdani, Hassan, Spiteller, Dieter, Popovic, Bojana, Spencer, Jonathan B., Staunton, James, Weissman, Kira J. and Leadlay, Peter F. (2005) Molecular basis of Celmer's rules: Stereochemistry of catalysis by isolated ketoreductase domains from modular polyketide synthases. Chemistry & Biology, 12 10: 1145-1153-1145-1153. doi:10.1016/j.chembiol.2005.08.017


Author Siskos, Alexandros P.
Baerga-Ortiz, Abel
Bali, Shilpa
Stein, Viktor
Mamdani, Hassan
Spiteller, Dieter
Popovic, Bojana
Spencer, Jonathan B.
Staunton, James
Weissman, Kira J.
Leadlay, Peter F.
Title Molecular basis of Celmer's rules: Stereochemistry of catalysis by isolated ketoreductase domains from modular polyketide synthases
Journal name Chemistry & Biology   Check publisher's open access policy
ISSN 1074-5521
1879-1301
Publication date 2005-10
Sub-type Article (original research)
DOI 10.1016/j.chembiol.2005.08.017
Volume 12
Issue 10
Start page 1145-1153
End page 1145-1153
Total pages 9
Place of publication London, U.K.
Publisher Cell Press
Language eng
Subject 0304 Medicinal and Biomolecular Chemistry
0305 Organic Chemistry
0601 Biochemistry and Cell Biology
Formatted abstract
A system is reported for the recombinant expression of individual ketoreductase (KR) domains from modular polyketide synthases (PKSs) and scrutiny of their intrinsic specificity and stereospecificity toward surrogate diketide substrates. The eryKR1 and the tylKR1 domains, derived from the first extension module of the erythromycin PKS and the tylosin PKS, respectively, both catalyzed reduction of (2R, S)-2-methyl-3-oxopentanoic acid N-acetylcysteamine thioester, with complete stereoselectivity and stereospecificity, even though the substrate is not tethered to an acyl carrier protein or an intact PKS multienzyme. In contrast, and to varying degrees, the isolated enzymes eryKR2, eryKR5, and eryKR6 exercised poorer control over substrate selection and the stereochemical course of ketoreduction. These data, together with modeling of diketide binding to KR1 and KR2, demonstrate the fine energetic balance between alternative modes of presentation of ketoacylthioester substrates to KR active sites.
©2005 Elsevier Ltd All rights reserved.
Keyword Binding sites
Catalysis
Kinetics
Models, molecular
Molecular conformation
Oxidoreductases
Polyketide synthases
Protein structure, tertiary
Stereoisomerism
Substrate specificity
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Institute for Molecular Bioscience - Publications
 
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Created: Wed, 24 Mar 2010, 10:16:34 EST by Jon Swabey on behalf of Institute for Molecular Bioscience