Cross-Linking/Mass Spectrometry Uncovers Details of Insulin-Like Growth Factor Interaction With Insect Insulin Binding Protein Imp-L2
Viola, Cristina M.
Brzozowski, Andrzej M.
Article (Published version)
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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 cross-linker 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 comple...x. 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.
Keywords:cross-linking / diazirine ring / IGF-1 / Imp-L2 / mass spectrometry
Source:Frontiers in Endocrinology, 2019, 10
- Frontiers Media S.A.
- Medical Research Council Grants MR/K000179/1 and MR/R009066/1
- Chemical biology for drugging undruggable targets (ChemBioDrug) (No. CZ.02.1.01/0.0/0.0/16_019/0000729)
- Czech Infrastructure for Integrative Structural Biology (LM2015043 CIISB for CMS BIOCEV funded by MEYS CR)
- Supplementary material: http://cherry.chem.bg.ac.rs/handle/123456789/3720