Fibrin Clot Architecture Regulated by Factor XIII

纤维蛋白凝块结构受因子 XIII 调节

• 批准号: 8574068
• 负责人: MURIEL C. MAURER
• 金额: $ 41.1万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-05 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8574068
• 关键词: Active Sites; Address; Amino Acids; Anticoagulants; Architecture; Binding; Biochemical; Biocompatible Materials; Biological Assay; Blood Clot; Blood coagulation; Cardiovascular Diseases; Cleaved cell; Coagulation Process; Complement; Cytolysis; Development; Distant; Environment; Enzymes; Factor XIII; Factor XIIIa; Fibrin; Fibrinogen; Fibrinopeptide A; Fibrinopeptide B; Genetic Polymorphism; Glutamine; Health; Hemorrhage; High Pressure Liquid Chromatography; Individual; Kinetics; Knowledge; Label; Lateral; Lead; MALDI-TOF Mass Spectrometry; Mass Spectrum Analysis; Measurement; Medical; Methods; Molecular; Multienzyme Complexes; Mutation; Network-based; P-2; Peptides; Physiological; Play; Positioning Attribute; Predisposition; Production; Property; Proteins; Research; Research Project Grants; Role; Secure; Site; Solutions; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Structure; Study models; TOCSY; Therapeutic; Thrombelastography; Thrombin; Time; Translating; Work; Wound Healing; base; crosslink; design; improved; mutant; novel therapeutics; polymerization; programs; public health relevance; stem

DESCRIPTION (provided by applicant): In blood coagulation, thrombin cleaves fibrinogen (AaBbg)2, sites for fibrin polymerization are revealed, and fibrin clot formation begins. Thrombin-activated Factor XIII is responsible for catalyzing the formation of covalent cross-links between fibrin molecules and in fibrin-enzyme complexes. The main objective of the research project is to examine how fibrin clot architecture can be regulated by FXIII. Intermolecular cross-links involving the fibrinogen aC region help secure lateral fibrin aggregation and lead to production of a more robust fibrin clot. Clot character can also be altered by influencing thrombin's ability to cleave the FXIII activation peptide segment. The roles of individual aC and FXIII amino acid regions in clot formation will be assessed. A greater understanding of these molecular details can be translated into new strategies to positively impact medical conditions. The vital biomaterial fibrin can be manipulated to create a stronger or weaker blood clot. FXIII and the resultant fibrin network are already known to play crucial roles in stemming blood loss, in wound healing, and in cardiovascular disease. For the development of new therapeutics to regulate blood clot structure, critical gaps in our knowledge must be resolved. More information is needed on 1) the kinetic and structural features of fibrinogen aC, a key substrate for FXIII and 2) the use of FXIII A2 mutants to fine tune FXIII activation and its subsequent functional effects. The proposed research for this application will thus address two hypotheses: 1) FXIII is hypothesized to take advantage of specific reactive glutamines within the ¿C (233-425) segment, along with local and more distant ¿C residues, to promote formation of fibrin ¿-¿ cross-links. Such covalent cross-links enhance intermolecular fibrin chain contacts, clot strength, and protection from degradation and 2) FXIII A2 V34X mutants are hypothesized to possess different abilities to be activated and the extent of tranglutaminase function can be used to alter fibrin clot character. The FXIII A2 mutants will include the common polymorphism L34 and the aromatic substitutions F34, W34, and Y34. The biochemical methods required to address these aims include mass spectrometry, solution NMR, kinetic assays, and measurements of fibrin clot formation and architecture. The knowledge gained from these studies will be used to improve the utilization of a biomaterials-based network for therapeutic purposes.

描述（由适用提供）：在血液凝结中，凝血蛋白切割纤维蛋白原（AABBG）2，纤维蛋白聚合的位点被发现，纤维蛋白凝块的形成开始。凝血酶激活的因子XIII负责催化纤维蛋白分子和纤维蛋白 - 酶复合物之间的共价交联。研究项目的主要目的是检查如何通过FXIII调节纤维蛋白血块结构。涉及纤维蛋白原AC区域的分子间交联有助于确保侧向纤维蛋白聚集并导致产生更健壮的纤维蛋白凝块。凝块特性也可以通过影响凝血酶清除FXIII激活肽段的能力来改变。将评估单个AC和FXIII氨基酸区域在凝块形成中的作用。对这些分子细节的更多了解可以转化为新的策略，以积极影响医疗状况。可以操纵重要的生物材料纤维蛋白，以产生更强或弱的血凝块。已知FXIII和由此产生的纤维蛋白网络在阻止失血，伤口愈合和心血管疾病中起着至关重要的作用。为了开发新的治疗剂以调节血凝块结构，必须解决我们知识的关键差距。需要有关1）纤维蛋白原AC的动力学和结构特征的更多信息，这是FXIII和FXIII的关键底物 2）使用FXIII A2突变体来微调FXIII激活及其随后的功能效应。 因此，针对本应用的拟议研究将解决两个假设：1）假设FXIII可以利用€c（233-425）节段内的特定反应性谷氨酰胺以及局部且较远的c残留物，以促进纤维蛋白的形成纤维蛋白。这种共价交联增强了分子间纤维蛋白链的接触，凝块强度和免受降解的保护，以及2）FXIII A2 V34X突变体被假设具有激活的潜在不同能力，并且可以使用Tranglutamiase功能的程度来改变Fibrin Clot特性。 FXIII A2突变体将包括常见的多态性L34和芳族取代F34，W34和Y34。解决这些目标所需的生化方法包括质谱，溶液NMR，动力学测定以及纤维蛋白凝块形成和结构的测量。从这些研究中获得的知识将用于改善基于生物材料的网络用于治疗目的的利用。