Sensitive and selective detection of free FXIII activation peptide: a potential marker of acute thrombotic events.
نویسندگان
چکیده
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 cleavage 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 detection. 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 monomers. We optimized our previously developed AP-FXIII ELISA by using 2 monoclonal antibodies. We determined high binding affinities between the antibodies and free AP-FXIII and demonstrated specific binding by epitope mapping analyses with surface plasmon resonance and enzyme-linked immunosorbent assay. Because 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 significantly from the rigid structure in the bound state. We suggest that the recognized epitope is either occluded in the noncleaved form or possesses a structure 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.
منابع مشابه
Sensitive and selective detection of free FXIII activation peptide: A potential marker of acute thrombotic events Short title: Selective detection of released AP-FXIII
متن کامل
THROMBOSIS AND HEMOSTASIS Sensitive and selective detection of free FXIII activation peptide: a potential marker of acute thrombotic events
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 cleavage 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 detection. We analyzed the st...
متن کاملFactor XIII activation peptide is released into plasma upon cleavage by thrombin and shows a different structure compared to its bound form.
The first step of coagulation factor XIII (FXIII) activation involves cleavage of the FXIII activation peptide (FXIII-AP) by thrombin. However, it is not known whether the FXIII-AP is released into plasma upon cleavage or remains attached to activated FXIII. The aim of the present work was to study the structure of free FXIII-AP, develop an assay for FXIII-AP determination in human plasma, and ...
متن کاملO10: Thrombo-Inflammation in Acute Ischemic Stroke
Ischemic stroke has been classified as a merely thrombotic disease, so the main goal of its treatment is the recanalization of the occluded vasculature. However, despite fast restoration of blood circulation, progressive stroke still develops in many patients, which has led to the concept of reperfusion injury. The underlying mechanism is only partly known. Though, it is accepted now, tha...
متن کاملPancreatic Differentiation of Sox 17 Knock-in Mouse Embryonic Stem Cells in Vitro
The way to overcome current limitations in the generation of glucose-responsive insulin-producing cells is selective enrichment of the number of definitive endoderm (DE) progenitor cells. Sox17 is the marker of mesendoderm and definitive endoderm. The aim of the present research was to study the potential of Sox17 knock-in CGR8 mouse embryonic stem (ES) cells to differentiate into insulin produ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Blood
دوره 115 24 شماره
صفحات -
تاریخ انتشار 2010