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Sensitive and selective detection of free FXIII activation peptide: a potential marker of acute thrombotic events


Ortner, E; Schroeder, V; Walser, R; Zerbe, O; Kohler, H-P (2010). Sensitive and selective detection of free FXIII activation peptide: a potential marker of acute thrombotic events. Blood, 115(24):5089-5096.

Abstract

Coagulation factor XIII (FXIII) stabilizes fibrin fibers and is therefore a major player in the maintenance of hemostasis. FXIII is activated by thrombin resulting in cleav- age and release of the FXIII activation peptide (AP-FXIII). The objective of this study was to characterize the released AP-FXIII and determine specific features that may be used for its specific detec- tion. We analyzed the structure of bound AP-FXIII within the FXIII A-subunit and interactions of AP-FXIII by hydrogen bonds with both FXIII A-subunit mono- mers. We optimized our previously devel- oped AP-FXIII ELISA by using 2 monoclo- nal antibodies. We determined high binding affinities between the antibodies and free AP-FXIII and demonstrated spe- cific binding by epitope mapping analy- ses with surface plasmon resonance and enzyme-linked immunosorbent assay. Be- cause the structure of free AP-FXIII had been characterized so far by molecular modeling only, we performed structural analysis by nuclear magnetic resonance. Recombinant AP-FXIII was largely flexible both in plasma and water, differing signifi- cantly from the rigid structure in the bound state. We suggest that the recog- nized epitope is either occluded in the noncleaved form or possesses a struc- ture that does not allow binding to the antibodies. On the basis of our findings, we propose AP-FXIII as a possible new marker for acute thrombotic events.

Abstract

Coagulation factor XIII (FXIII) stabilizes fibrin fibers and is therefore a major player in the maintenance of hemostasis. FXIII is activated by thrombin resulting in cleav- age and release of the FXIII activation peptide (AP-FXIII). The objective of this study was to characterize the released AP-FXIII and determine specific features that may be used for its specific detec- tion. We analyzed the structure of bound AP-FXIII within the FXIII A-subunit and interactions of AP-FXIII by hydrogen bonds with both FXIII A-subunit mono- mers. We optimized our previously devel- oped AP-FXIII ELISA by using 2 monoclo- nal antibodies. We determined high binding affinities between the antibodies and free AP-FXIII and demonstrated spe- cific binding by epitope mapping analy- ses with surface plasmon resonance and enzyme-linked immunosorbent assay. Be- cause the structure of free AP-FXIII had been characterized so far by molecular modeling only, we performed structural analysis by nuclear magnetic resonance. Recombinant AP-FXIII was largely flexible both in plasma and water, differing signifi- cantly from the rigid structure in the bound state. We suggest that the recog- nized epitope is either occluded in the noncleaved form or possesses a struc- ture that does not allow binding to the antibodies. On the basis of our findings, we propose AP-FXIII as a possible new marker for acute thrombotic events.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Uncontrolled Keywords:stroke thrombosis antibody
Date:June 2010
Deposited On:27 Oct 2010 12:46
Last Modified:05 Apr 2016 14:14
Publisher:American Society of Hematology
ISSN:0006-4971
Additional Information:This research was originally published in Blood. Blood, 17 June 2010, Vol. 115, No. 24, pp. 5089-5096. Copyright by the American Society of Hematology.
Publisher DOI:https://doi.org/10.1182/blood-2009-11-253062
PubMed ID:20375315

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