Coagulation Factor XIIIa Undergoes a Conformational Change Evoked by Glutamine Substrate

ECU Author/Contributor (non-ECU co-authors, if there are any, appear on document)

Paul D. Bishop (Creator)

Olga Mitkevich (Creator)

Gennady P. Samokhin (Creator)

John R. Shainoff (Creator)

Gary B. Smejkal (Creator)

David C. Teller (Creator)

Vivian Yee (Creator)

Institution

East Carolina University (ECU )

Web Site: http://www.ecu.edu/lib/

Abstract: Coagulation factor XIIIa, plasma transglutaminase(endo-g-glutamine:e-lysine transferase EC 2.3.2.13) cata-lyzes isopeptide bond formation between glutamine andlysine residues and rapidly cross-links fibrin clots. Amonoclonal antibody (5A2) directed to a fibrinogen Aa-chain segment 529 –539 was previously observed fromanalysis of end-stage plasma clots to block fibrina-chaincross-linking. This prompted the study of its effect onnonfibrinogen substrates, with the prospect that 5A2was inhibiting XIIIa directly. It inhibited XIIIa-cata-lyzed incorporation of the amine donor substrate dan-sylcadaverine into the glutamine acceptor dimethylca-sein in an uncompetitive manner with respect todimethylcasein utilization and competitively with re-spect to dansylcadaverine. Uncompetitive inhibitionwas also observed with the synthetic glutamine sub-strate, LGPGQSKVIG. Theoretically, uncompetitive in-hibition arises from preferential interaction of the in-hibitor with the enzyme-substrate complex but is alsofound to inhibitg-chain cross-linking. The conjunctionof the uncompetitive and competitive modes of inhibi-tion indicates in theory that this bireactant system in-volves an ordered reaction in which docking of the glu-tamine substrate precedes the amine exchange. Thepresence of substrate enhanced binding of 5A2 to XIIIa,an interaction deemed to occur through a C-terminalsegment of the XIIIa A-chain (643– 658,GSDMTVTVQFT-NPLKE), 55% of which comprises sequences occurring inthe fibrinogen epitope Aa-(529 –540) (GSESGIFTNTKE).Removal of the C-terminal domain from XIIIa abolishesthe inhibitory effect of 5A2 on activity. Crystallographicstudies on recombinant XIIIa place the segment 643– 658in the region of the groove through which glutaminesubstrates access the active site and have predicted thatfor catalysis, a conformational change may accompanyglutamine-substrate binding. The uncompetitive inhibi-tion and the substrate-dependent binding of 5A2 pro-vide evidence for the conformational change.

Additional Information

Publication

Other

Language: English

Date: 2023

Subjects

coagulation factor glutamine clots

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