Hydroxyl radical footprinting analysis of a human haptoglobin-hemoglobin complex

0553501 - MBÚ 2023 RIV NL eng J - Journal Article
Loginov, Dmitry Sergej - Fiala, Jan - Brechlin, P. - Kruppa, G. - Novák, Petr
Hydroxyl radical footprinting analysis of a human haptoglobin-hemoglobin complex.
Biochimica Et Biophysica Acta-Proteins and Proteomics. Roč. 1870, č. 2 (2022), č. článku 140735. ISSN 1570-9639. E-ISSN 1878-1454
R&D Projects: GA ČR(CZ) GA19-16084S; GA MŠMT(CZ) LQ1604
EU Projects: European Commission(XE) 823839 - EPIC-XS
Institutional support: RVO:61388971
Keywords : fast photochemical oxidation * identification * proteins * fpop * Footprinting * Haptoglobin * Hemoglobin * timsTOF pro
OECD category: Biochemistry and molecular biology
Impact factor: 3.2, year: 2022
Method of publishing: Open access
https://www.sciencedirect.com/science/article/pii/S1570963921001412?via%3Dihub

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 Hba, and Tyr280 and Trp284 in Hp beta. 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 beta Trp37 and Hp beta Phe292 was not confirmed by our data. Data are available via ProteomeXchange with identifier PXD021621.
Permanent Link: http://hdl.handle.net/11104/0330400