Cross-Linking/Mass Spectrometry Uncovers Details of Insulin-Like Growth Factor Interaction With Insect Insulin Binding Protein Imp-L2

0518634 - MBÚ 2020 RIV CH eng J - Journal Article
Pompach, Petr - Viola, C. M. - Radosavljević, Jelena - Lin, Jingjing - Jiráček, Jiří - Brzozowski, A. M. - Selicharová, Irena
Cross-Linking/Mass Spectrometry Uncovers Details of Insulin-Like Growth Factor Interaction With Insect Insulin Binding Protein Imp-L2.
Frontiers in Endocrinology. Roč. 10, OCT 9 2019 (2019), č. článku 695. ISSN 1664-2392. E-ISSN 1664-2392
R&D Projects: GA MŠMT(CZ) EF16_019/0000729
Institutional support: RVO:61388971 ; RVO:61388963
Keywords : IGF-1 * diazirine ring * mass spectrometry
OECD category: Biochemistry and molecular biology; Biochemical research methods (UOCHB-X)
Impact factor: 3.644, year: 2019
Method of publishing: Open access
https://www.frontiersin.org/articles/10.3389/fendo.2019.00695/full

Structural details of changes accompanying interaction between insulin-related hormones and their binding partners are often enigmatic. Here, cross-linking/mass spectrometry could complement structural techniques and reveal details of these protein-protein interfaces. We used such approach to clarify missing structural description of the interface in human insulin-like growth factor (IGF-1): Drosophila melanogaster imaginal morphogenesis protein-late 2 protein (Imp-L2) complex which we studied previously by X-ray crystallography. We crosslinked these proteins by heterobifunctional crosslinker sulfosuccinimidyl 4,4'-azidopentanoate (Sulfo-SDA) for the subsequent mass spectrometry (MS) analysis. The MS analysis revealed IGF-1:Imp-L2 interactions which were not resolved in the crystal structure of this assembly, and they converged with X-ray results, indicating the importance of the IGF-1 N-terminus interaction with the C-terminal (185-242) part of the Imp-L2 for stability of this complex. Here, we also showed the advantage and reliability of MS approach in solving details of protein-protein interactions that are too flexible for solid state structural methods.
Permanent Link: http://hdl.handle.net/11104/0303739