Document Detail


Elimination of P1 arginine 393 interaction with underlying glutamic acid 255 partially activates antithrombin III for thrombin inhibition but not factor Xa inhibition.
MedLine Citation:
PMID:  11971909     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The mechanism for heparin activation of antithrombin III has been postulated to involve disruption of interactions between its reactive loop P1 residue and Glu(255) on the underlying protein surface. To test this hypothesis, the potential P1-constraining Arg(393)-Glu(255) hydrogen bond and ionic interactions were eliminated by converting Glu(255) to alanine. E255A and wild-type ATIIIs have identical reactive loop sequences (including the P1 and P14 residues), but differ in that Glu(255)-mediated, P1-constraining interactions with the underlying surface cannot form in the mutant. Relative to its wild-type parent, E255A had a 5-fold higher affinity for heparin and pentasaccharide. In the absence of cofactor, E255A exhibited a 5-fold activation of thrombin inhibition but no activation of factor Xa inhibition. Pentasaccharide addition elicited no further activation of thrombin inhibition but increased the factor Xa inhibition rate 100-fold. E255A heparin-dependent thrombin and factor Xa inhibition rates were 1000- and 2-fold faster, respectively, than pentasaccharide-catalyzed rates. Although "approximation" is the predominant factor in heparin activation of ATIII thrombin inhibition, and removal of the P1 constraint plays a distinct but minor role, the primary determinant for activation of factor Xa inhibition is the pentasaccharide-induced conformational change, with approximation making a further minor contribution, and removal of the P1 constraint playing no role at all.
Authors:
Mohamad Aman Jairajpuri; Aiqin Lu; Susan C Bock
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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.     Date:  2002-04-23
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  277     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2002 Jul 
Date Detail:
Created Date:  2002-07-01     Completed Date:  2002-08-27     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  24460-5     Citation Subset:  IM    
Affiliation:
Department of Medicine, University of Utah, Salt Lake City, Utah 84132, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Amino Acid Substitution
Animals
Antithrombin III / chemistry,  metabolism*
Arginine*
Base Sequence
Binding Sites
Cell Line
Factor Xa / metabolism*
Glutamic Acid*
Heparin / pharmacology
Humans
Models, Molecular
Mutagenesis, Site-Directed
Polymerase Chain Reaction
Protein Conformation
Recombinant Proteins / chemistry,  metabolism
Spodoptera
Thrombin / antagonists & inhibitors*
Transfection
Grant Support
ID/Acronym/Agency:
R01-HL30712/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Recombinant Proteins; 56-86-0/Glutamic Acid; 74-79-3/Arginine; 9000-94-6/Antithrombin III; 9005-49-6/Heparin; EC 3.4.21.5/Thrombin; EC 3.4.21.6/Factor Xa

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


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