Document Detail

Breaking the covalent connection: Chain connectivity and the catalytic reaction of PMM/PGM.
MedLine Citation:
PMID:  20512975     Owner:  NLM     Status:  MEDLINE    
Fragment complementation has been used to investigate the role of chain connectivity in the catalytic reaction of phosphomannomutase/phosphoglucomutase (PMM/PGM) from Pseudomonas aeruginosa, a human pathogen. A heterodimer of PMM/PGM, created from fragments corresponding to its first three and fourth domains, was constructed and enzyme activity reconstituted. NMR spectra demonstrate that the fragment corresponding to the fourth (C-terminal) domain exists as a highly structured, independent folding domain, consistent with its varied conformation observed in enzyme-substrate complexes. Steady-state kinetics and thermodynamics studies reported here show that complete conformational freedom of Domain 4, because of the break in the polypeptide chain, is deleterious to catalytic efficiency primarily as a consequence of increased entropy. This extends observations from studies of the intact enzyme, which showed that the degree of flexibility of a hinge region is controlled by the precise sequence of amino acids optimized through evolutionary constraints. This work also sheds light on the functional advantage gained by combining separate folding domains into a single polypeptide chain.
Andrew M Schramm; Dale Karr; Ritcha Mehra-Chaudhary; Steven R Van Doren; Cristina M Furdui; Lesa J Beamer
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Protein science : a publication of the Protein Society     Volume:  19     ISSN:  1469-896X     ISO Abbreviation:  Protein Sci.     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-05-31     Completed Date:  2010-09-02     Revised Date:  2013-05-29    
Medline Journal Info:
Nlm Unique ID:  9211750     Medline TA:  Protein Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1235-42     Citation Subset:  IM    
Department of Biochemistry, University of Missouri, Columbia, Missouri 65211, USA.
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MeSH Terms
Bacterial Proteins / chemistry,  genetics,  metabolism*
Escherichia coli / genetics
Models, Molecular
Multiprotein Complexes / chemistry,  genetics,  metabolism*
Nuclear Magnetic Resonance, Biomolecular
Phosphoglucomutase / chemistry,  genetics,  metabolism*
Phosphotransferases (Phosphomutases) / chemistry,  genetics,  metabolism*
Protein Structure, Tertiary
Pseudomonas aeruginosa / enzymology,  genetics
Recombinant Proteins / chemistry,  genetics,  metabolism
Grant Support
5R03 AI074779-02/AI/NIAID NIH HHS; R03 AI074779-01/AI/NIAID NIH HHS; R03 AI074779-02/AI/NIAID NIH HHS
Reg. No./Substance:
0/Bacterial Proteins; 0/Multiprotein Complexes; 0/Recombinant Proteins; EC 5.4.2.-/Phosphotransferases (Phosphomutases); EC; EC

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

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