Document Detail

Expression, purification, and structural analysis of the trimeric form of the catalytic domain of the Escherichia coli dihydrolipoamide succinyltransferase.
MedLine Citation:
PMID:  10739245     Owner:  NLM     Status:  MEDLINE    
The dihydrolipoamide succinyltransferase (E2o) component of the alpha-ketoglutarate dehydrogenase complex catalyzes the transfer of a succinyl group from the S-succinyldihydrolipoyl moiety to coenzyme A. E2o is normally a 24-mer, but is found as a trimer when E2o is expressed with a C-terminal [His]6 tag. The crystal structure of the trimeric form of the catalytic domain (CD) of the Escherichia coli E2o has been solved to 3.0 A resolution using the Molecular Replacement method. The refined model contains an intact trimer in the asymmetric unit and has an R-factor of 0.257 (Rfree = 0.286) for 18,699 reflections between 10.0 and 3.0 A resolution. The core of tE2oCD (residues 187-396) superimposes onto that of the cubic E2oCD with an RMS difference of 0.4 A for all main-chain atoms. The C-terminal end of tE2oCD (residues 397-404) rotates by an average of 37 degrees compared to cubic E2oCD, disrupting the normal twofold interface. Despite the alteration of quaternary structure, the active site of tE2oCD shows no significant differences from that of the cubic E2oCD, although several side chains in the active site are more ordered in the trimeric form of E2oCD. Analysis of the available sequence data suggests that the majority of E2 components have active sites that resemble that of E. coli E2oCD. The remaining E2 components can be divided into three groups based on active-site sequence similarity. Analysis of the surface properties of both crystal forms of E. coli E2oCD suggests key residues that may be involved in the protein-protein contacts that occur between the catalytic and lipoyl domains of E2o.
J E Knapp; D Carroll; J E Lawson; S R Ernst; L J Reed; M L Hackert
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Protein science : a publication of the Protein Society     Volume:  9     ISSN:  0961-8368     ISO Abbreviation:  Protein Sci.     Publication Date:  2000 Jan 
Date Detail:
Created Date:  2000-05-05     Completed Date:  2000-05-05     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  9211750     Medline TA:  Protein Sci     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  37-48     Citation Subset:  IM    
Department of Chemistry and Biochemistry, The University of Texas at Austin, 78712, USA.
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MeSH Terms
Acyltransferases / chemistry*,  isolation & purification
Amino Acid Sequence
Binding Sites
Catalytic Domain
Crystallography, X-Ray
Escherichia coli / chemistry*
Models, Molecular
Molecular Sequence Data
Protein Structure, Quaternary
Grant Support
Reg. No./Substance:
EC 2.3.-/Acyltransferases; EC succinyltransferase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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