Factor XIII Activation and Substrate Specificity

XIII 因子激活和底物特异性

基本信息

批准号：
7849591
负责人：
MURIEL C. MAURER
金额：
$ 24.83万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-07-08 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7849591
关键词：
Active Sites Address Adopted Amides Amino Acid Substitution Amino Acids Antiplasmin Architecture Arteriosclerosis Binding Binding Sites Biological Assay Biological Models Blood Blood Clot Blood coagulation Calcium Catalytic Domain Chemicals Cleaved cell Coagulants Coagulation Process Complement Complex Deuterium Docking Enzymes Event Factor XIII Factor XIIIa Fibrin Fibrinogen Genetic Polymorphism Glutamine High Pressure Liquid Chromatography Hydrogen Hydrolysis Individual Kinetics Lead Ligand Binding MALDI-TOF Mass Spectrometry Maps Medical Methods Modification Molecular Molecular Target Monitor Multienzyme Complexes Myocardial Infarction N-terminal NOESY Peptides Plasminogen Activator Inhibitor 2 Positioning Attribute Property Proteins Reaction Relative (related person)Reliance Research Research Personnel Research Project Grants Roentgen Rays Role Series Site Sodium Solutions Solvents Stroke Structure Substrate Specificity Surface Surface Plasmon Resonance Testing Thrombin Time Transglutaminases Vitronectin Wound Healing base clotting enzyme crosslink design experience extracellular flexibility heart disease risk inhibitor/antagonist mutant novel polymerization programs

项目摘要

DESCRIPTION (provided by applicant): In blood clotting, the enzyme thrombin cleaves fibrinogen, sites for fibrin polymerization are revealed, and fibrin clot formation begins. Activated Factor XIII is responsible for catalyzing the formation of covalent crosslinks between fibrin molecules and in fibrin-enzyme complexes. Factor XIII can be activated through cleavage of an activation peptide segment by thrombin or nonproteolytically in the presence of high calcium or high sodium/low calcium. The long-term objective of our research project is to examine in solution the structural features that govern the activation and substrate specificity of Factor XIII. Understanding these molecular details is critical considering the role of FXIII in increasing the risk of heart disease, stroke, and arteriosclerosis. The specific aims are an extension of ongoing studies and will address three hypotheses: 1) The reactive glutamines of certain FXIII substrates primarily target a distinctive surface within the transglutaminase active site region. Such studies are important since the substrate specificity of FXIII is not clearly defined, 2) FXIII undergoes subtle conformational changes upon activation. The degree and surface coverage of these changes will increase following introduction of a substrate or inhibitor, and 3) The FXIII activation peptide segment utilizes key positions to promote effective interactions with a thrombin surface whose properties are regulated by individual thrombin residues. A model system for such studies is FXIII V34L, a common polymorphism that has been correlated with protection against myocardial infarction and is more susceptible to thrombin cleavage than the native sequence. To test these hypotheses, a combination of kinetic studies, hydrogen/deuterium exchange, chemical modification methods, MALDI-TOF mass spectrometry, solution NMR, and surface plasmon resonance methods will be used. Information will be obtained on the selectivity of FXIII for glutamine-containing substrates, on the solvent accessibility changes that FXIII undergoes upon activation and ligand binding, and on the kinetic/structural features of the thrombin - FXIII Activation Peptide complex. Lay Summary: Factor XIII is a key enzyme in the formation of blood clots. While very important for wound healing, the clots can also increase risk for heart disease and stroke. Research will focus on understanding how Factor XIII is turned on and how it selects its targets. Studies may lead to novel medical strategies to control the actions of Factor XIII and the resultant clot architecture.

描述（由申请人提供）：在血液凝结中，酶凝血酶裂解纤维蛋白原，纤维蛋白聚合的位点，并开始纤维蛋白凝块的形成。活化因子XIII负责催化纤维蛋白分子和纤维蛋白 - 酶复合物之间共价交联的形成。在存在高钙或高钠/低钙的情况下，可以通过凝血酶或非蛋白溶液通过凝血酶或非蛋白质来激活XIII。我们研究项目的长期目标是在解决方案中检查控制因子XIII的激活和底物特异性的结构特征。考虑到FXIII在增加心脏病，中风和动脉硬化的风险中的作用，了解这些分子细节至关重要。具体目的是正在进行的研究的扩展，并将解决三个假设：1）某些FXIII底物的反应性谷氨酰胺主要针对跨谷氨酰胺酶活性位点区域内的独特表面。此类研究很重要，因为未明确定义FXIII的底物特异性，2）FXIII在激活时经历细微的构象变化。引入底物或抑制剂后，这些变化的程度和表面覆盖率将增加，3）FXIII激活肽段利用关键位置来促进与凝血酶表面的有效相互作用，其特性受单个凝血酶残基调节。用于此类研究的模型系统是FXIII V34L，这是一种常见的多态性，与对心肌梗死的保护相关，并且比天然序列更容易受到凝血酶裂解的影响。为了检验这些假设，将使用动力学研究，氢/氘交换，化学修饰方法，MALDI-TOF质谱法，溶液NMR和表面等离子体共振方法的组合。将获得有关FXIII对含谷氨酰胺底物的选择性的信息，FXIII在激活和配体结合后经历的溶剂可访问性变化以及凝血酶-FXIII激活肽复合物的动力学/结构特征。摘要摘要：XIII因子是血块形成的关键酶。尽管对于伤口愈合非常重要，但凝块也可以增加心脏病和中风的风险。研究将集中于了解XIII的打开方式以及其如何选择目标。研究可能会导致新的医疗策略来控制第十三因子的作用和最终的凝块结构。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mapping of factor XIII solvent accessibility as a function of activation state using chemical modification methods.

使用化学修饰方法将因子 XIII 溶剂可及性映射为激活状态的函数。

DOI：
10.1021/bi049260
发表时间：
2004
期刊：
Biochemistry.
影响因子：
0
作者：
TurnerJr,BrianT;Sabo,TMichael;Wilding,Diana;Maurer,MurielC
通讯作者：
Maurer,MurielC

A non-reactive glutamine residue of alpha2-antiplasmin promotes interactions with the factor XIII active site region.

DOI：
10.1111/j.1538-7836.2009.03583.x
发表时间：
2009-11
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
0
作者：
Cleary DB;Doiphode PG;Sabo TM;Maurer MC
通讯作者：
Maurer MC

Ranking reactive glutamines in the fibrinogen αC region that are targeted by blood coagulant factor XIII.

对凝血因子 XIII 所针对的纤维蛋白原 αC 区域中的反应性谷氨酰胺进行排名。

DOI：
10.1182/blood-2015-09-672303
发表时间：
2016
期刊：
Blood
影响因子：
20.3
作者：
Mouapi,KellyNjine;Bell,JacobD;Smith,KerrieA;Ariëns,RobertAS;Philippou,Helen;Maurer,MurielC
通讯作者：
Maurer,MurielC

Deciphering Conformational Changes Associated with the Maturation of Thrombin Anion Binding Exosite I.