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Blood, 1 June 2004, Vol. 103, No. 11, pp. 4157-4163. Prepublished online as a Blood First Edition Paper on February 5, 2004; DOI 10.1182/blood-2003-12-4296. This Article Full Text (PDF) All Versions of this Article: 2003-12-4296v1 103/11/4157    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Okumura, N. Articles by Lord, S. T Search for Related Content PubMed PubMed Citation Articles by Okumura, N. Articles by Lord, S. T Social Bookmarking                   What's this? Submitted December 19, 2003 Accepted January 26, 2004 Substitution of the -chain Asn 308 disturbs the D:D interface affecting fibrin polymerization, fibrinopeptide B release, and FXIII catalyzed cross-linking Nobuo Okumura*, Oleg V Gorkun, Fumiko Terasawa, and Susan T Lord Biomedical Laboratory Sciences, School of Health Sciences, Shinshu University, Matsumoto, Nagano, Japan Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA * Corresponding author; email: nobuoku{at}gipac.shinshu-u.ac.jp. Crystallographic structures indicate that -chain residue Asn308 participates in D:D interactions and indeed substitutions of Asn308 with Lys or Ile have been identified in dysfibrinogens with impaired polymerization. To probe the role of Asn308 in polymerization, we synthesized three variant fibrinogens: Asn308 changed to Lys (N308K), Ile (N308I), and Ala (N308A). We measured thrombin-catalyzed polymerization by turbidity, fibrinopeptide release by HPLC, and Factor XIIIa-catalyzed cross-linking by SDS-PAGE. In the absence of added calcium, polymerization was clearly impaired with all three variants. In contrast, at 0.1 mM calcium, only polymerization of N308K remained markedly abnormal. Thrombin-catalyzed fibrinopeptide B (FpB) release was delayed in the absence of calcium, while at 1 mM calcium FpB release was delayed only with N308K. Factor XIIIa-catalyzed - dimer formation was delayed with fibrinogen (in absence of thrombin) while with fibrin (in presence of thrombin) - dimer formation of only N308K was delayed. These data corroborate the recognized link between FpB release and polymerization. They show fibrin cross-link formation likely depends on the structure of protofibrils. Together, our results show substitution of Asn308 with a hydrophobic residue altered neither polymer formation nor polymer structure at physiologic calcium concentrations, while substitution with lysine altered both. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: T. A. Dugan, V. W.-C. Yang, D. J. McQuillan, and M. Hook Decorin Modulates Fibrin Assembly and Structure J. Biol. Chem., December 15, 2006; 281(50): 38208 - 38216. [Abstract] [Full Text] [PDF] S. Kani, F. Terasawa, K. Yamauchi, M. Tozuka, and N. Okumura Analysis of fibrinogen variants at {gamma}387Ile shows that the side chain of {gamma}387 and the tertiary structure of the {gamma}C-terminal tail are important not only for assembly and secretion of fibrinogen but also for lateral aggregation of protofibrils and XIIIa-catalyzed {gamma}-{gamma} dimer formation Blood, September 15, 2006; 108(6): 1887 - 1894. [Abstract] [Full Text] [PDF]

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