BioCeV - Institute of Microbiology of the CAS, Prumyslova 595, CZ-252 50 Vestec, Czech Republic; Orekhovich Institute of Biomedical Chemistry, Pogodinskaja str. 10, 119191 Moscow, Russia.
BioCeV - Institute of Microbiology of the CAS, Prumyslova 595, CZ-252 50 Vestec, Czech Republic; Faculty of Science, Charles University, Hlavova 8, CZ-128 20 Prague, Czech Republic.
Biochim Biophys Acta Proteins Proteom. 2022 Feb 1;1870(2):140735. doi: 10.1016/j.bbapap.2021.140735. Epub 2021 Nov 3.
Methods of structural mass spectrometry have become more popular to study protein structure and dynamics. Among them, fast photochemical oxidation of proteins (FPOP) has several advantages such as irreversibility of modifications and more facile determination of the site of modification with single residue resolution. In the present study, FPOP analysis was applied to study the hemoglobin (Hb) - haptoglobin (Hp) complex allowing identification of respective regions altered upon the complex formation. FPOP footprinting using a timsTOF Pro mass spectrometer revealed structural information for 84 and 76 residues in Hp and Hb, respectively, including statistically significant differences in the modification extent below 0.3%. The most affected residues upon complex formation were Met76 and Tyr140 in Hbα, and Tyr280 and Trp284 in Hpβ. The data allowed determination of amino acids directly involved in Hb - Hp interactions and those located outside of the interaction interface yet affected by the complex formation. Also, previously modeled interaction between Hb βTrp37 and Hp βPhe292 was not confirmed by our data. Data are available via ProteomeXchange with identifier PXD021621.
结构质谱学方法已越来越多地用于研究蛋白质结构和动力学。其中，快速光化学氧化蛋白质（FPOP）具有几个优点，例如修饰的不可逆性以及更易于确定修饰的位置，具有单个残基分辨率。在本研究中，应用 FPOP 分析研究血红蛋白（Hb）-触珠蛋白（Hp）复合物，允许鉴定复合物形成时各自发生变化的区域。使用 timsTOF Pro 质谱仪进行 FPOP 足迹分析分别揭示了 Hp 和 Hb 中 84 个和 76 个残基的结构信息，包括修饰程度的统计学显着差异低于 0.3%。在复合物形成过程中受影响最大的残基是 Hbα中的 Met76 和 Tyr140，以及 Hpβ中的 Tyr280 和 Trp284。该数据允许确定直接参与 Hb-Hp 相互作用的氨基酸以及位于相互作用界面之外但受复合物形成影响的氨基酸。此外，我们的数据并未证实先前建模的 HbβTrp37 和 HpβPhe292 之间的相互作用。数据可通过 ProteomeXchange 以标识符 PXD021621 获得。
