Chowdhury Abu Sayeed, Oxford Julia Thom
Biomolecular Sciences Graduate Program, Boise State University, 1910 University Drive, Boise, Idaho 83725, United States.
Biomolecular Research Institute, Boise State University, 1910 University Drive, Boise, Idaho 83725, United States.
Biochemistry. 2025 Feb 4;64(3):735-747. doi: 10.1021/acs.biochem.4c00434. Epub 2025 Jan 22.
The amino-terminal domain of collagen α1(XI) plays a key role in controlling fibrillogenesis. However, the specific mechanisms through which various isoforms of collagen α1(XI) regulate this process are not fully understood. We measured the kinetics of collagen type I self-assembly in the presence of specific collagen α1(XI) isoforms. Molecular dynamics simulations, protein-protein docking studies, and molecular mechanics Poisson-Boltzmann surface area were utilized to understand the molecular mechanisms. In vitro, in silico, and thermodynamic studies demonstrated an isoform-specific effect on self-assembly kinetics. Our results indicate isoform-specific differences in the rate constants, activation energy, and free energy of binding. These differences may result from isoform-specific interaction dynamics and modulation of steric hindrance due to the chemically distinct variable regions. We show that isoform A interacts with collagen type I due in part to the acidic variable region, increasing the activation energy of fibril growth while decreasing the rate constant during the growth phase. In contrast, the basic variable region of isoform B may result in less steric hindrance than isoform A. Isoform 0 demonstrated the highest activation energy and the lowest rate constant during the growth phase. Although the presence of isoforms reduced the rate constants for fibril growth, an increase in total turbidity during the plateau phase was observed compared to controls. Overall, these results are consistent with collagen α1(XI) NTD isoforms facilitating fibrillogenesis by increasing the final yield by reducing the rate of the lag and/or growth phases, while extending the duration of the growth phase.
胶原蛋白α1(XI)的氨基末端结构域在控制纤维形成过程中起关键作用。然而,胶原蛋白α1(XI)的各种异构体调节这一过程的具体机制尚未完全了解。我们测量了在特定胶原蛋白α1(XI)异构体存在下I型胶原蛋白自组装的动力学。利用分子动力学模拟、蛋白质-蛋白质对接研究和分子力学泊松-玻尔兹曼表面积来理解分子机制。体外、计算机模拟和热力学研究表明,异构体对自组装动力学具有特异性影响。我们的结果表明,在速率常数、活化能和结合自由能方面存在异构体特异性差异。这些差异可能源于异构体特异性相互作用动力学以及由于化学性质不同的可变区对空间位阻的调节。我们发现异构体A与I型胶原蛋白相互作用,部分原因是其酸性可变区,这增加了纤维生长的活化能,同时降低了生长阶段的速率常数。相比之下,异构体B的碱性可变区可能导致比异构体A更小的空间位阻。异构体0在生长阶段表现出最高的活化能和最低的速率常数。尽管异构体的存在降低了纤维生长的速率常数,但与对照组相比,在平台期观察到总浊度增加。总体而言,这些结果与胶原蛋白α1(XI) NTD异构体通过降低延迟和/或生长阶段的速率来提高最终产量,同时延长生长阶段的持续时间从而促进纤维形成是一致的。