Mapping protein structural changes by quantitative cross-linking

0455585 - MBÚ 2016 RIV US eng J - Článek v odborném periodiku
Kukačka, Zdeněk - Strohalm, Martin - Kavan, Daniel - Novák, Petr
Mapping protein structural changes by quantitative cross-linking.
Methods. Roč. 89, NOV 2015 (2015), s. 112-120. ISSN 1046-2023. E-ISSN 1095-9130
Grant CEP: GA MŠMT(CZ) EE2.3.20.0055; GA MŠMT(CZ) EE2.3.30.0003; GA MŠMT(CZ) ED1.1.00/02.0109
Grant ostatní: OPPC(XE) CZ.2.16/3.1.00/24023
Institucionální podpora: RVO:61388971
Klíčová slova: Chemical cross-linking * Proteolysis * Mass spectrometry
Kód oboru RIV: CE - Biochemie
Impakt faktor: 3.503, rok: 2015

Chemical cross-linking is a promising technology for protein tertiary structure determination. Though the data has low spatial resolution, it is possible to obtain it at physiological conditions on proteins that are not amenable to standard high resolution techniques such as X-ray, NMR analysis and cryo-EM. Here we demonstrate the utilization of isotopically labeled chemical cross-linking to visualize protein conformation rearrangements. Since calmodulin exists in two distinct conformations (calcium-free and calcium-containing forms), we selected this protein for testing the potential and the limits of a new technique. After cross-linking of both calmodulin forms, the calcium-free and calcium-containing forms were mixed together and digested under different conditions and the products of proteolysis were monitored using high resolution mass spectrometry. Finally, the ratios of heavy/light cross-links were calculated by mMass open source platform.
Trvalý link: http://hdl.handle.net/11104/0256211