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鉴定XIII因子β-三明治残留物介导谷氨酸基质结合和激活裂变

Rameesa D Syed Mohammed¹, Kellianne M Piell², Muriel C Maurer¹

¹Department of Chemistry, University of Louisville, Louisville, Kentucky, United States.

Thrombosis and haemostasis

|December 1, 2023

概括

因子XIII (FXIII) A亚单位残留物K156,F157,R158,R171和R174对于结合纤维素和活性素等生理基质至关重要. 这些残留物也会影响血栓激活,影响交叉连接能力.

科学领域:

生物化学生物化学
分子生物学分子生物学
蛋白质科学蛋白质科学

背景情况:

因子XIII (FXIII) 对于血液静止,伤口愈合和通过交叉连接基质的骨代谢至关重要.
FXIII活性与静脉血栓塞栓症 (VTE) 有关,是药物点.
了解FXIII-基质相互作用对于药物开发至关重要,之前的工作涉及FXIII-A子单元的β-三明治域在基质识别中.

研究的目的:

在负责基质识别的FXIII-A子单位内识别特定的残留物.
调查这些残留物在FXIII激活和交联活动中的作用.
阐明了FXIII-基质相互作用背后的分子机制.

主要方法:

再组合的FXIII-A蛋白质变体的生成 (WT,K156E,F157L,R158Q/E,R171Q,R174E).
对蛋白质溶解 (FXIII-A*) 和非蛋白质溶解 (FXIII-A°) 形式进行激活和交叉链接活动的分析.
使用SDS-PAGE和MALDI-TOFMS来评估生理基质 (Fbg αC,纤维素,活性蛋白) 和基质的交联.

主要成果:

与WT相比,所有FXIII-A*变体都显示出纤维素和动素的交叉连接减少.
FXIII-A* WT表现出高于FXIII-A°的活性,表明活性形式的基质结合部位暴露.
变种R158Q/E,R171Q和R174E显示Fbg αC交叉连接显著下降,接近FXIII-A°水平.
通过WT和变体对基质的类似交叉链接表明识别部位与催化部位不同.
FXIII-A R158E和R171Q变种表现出较慢的血栓激活,可能是由于与激活的H结合中断.

结论:

在FXIII-A中的K156,F157,R158,R171和R174残留物是纤维素和活性蛋白的结合部位的组成部分.
在激素激活过程中,R158和R171在调节激活裂变方面发挥着关键作用.
这些发现为FXIII基质相互作用及其对交叉连接功能的影响提供了新的分子见解.

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