Enabling Photoactivated Cross-Linking Mass Spectrometric Analysis of Protein Complexes by Novel MS-Cleavable Cross-Linkers
Craig Gutierrez, Leah Salituro, Clinton Yu, Xiaorong Wang, Sadie F. DePeter, Scott D. Rychnovsky, Lan Huang
Abstract
Although photochemistry complements residue-specific chemistry through labeling amino acids nonspecifically, existing photo-cross-linking reagents are thus far inapplicable to multisubunit protein complexes owing to low yields and high complexities of photo-cross-linked products. The development of the three sulfoxidecontaining MS-cleavable photoreactive SDASO cross-linkers permits MSn-based analytical workflow for accurate identification of photo-cross-linked peptides, enabling complex PPI profiling for the first time. This work has established a solid foundation for future applications of photo-crosslinking in complex XL-MS studies.
Topics & Concepts
ChemistryDiazirineMass spectrometryCombinatorial chemistryResidue (chemistry)Protein–protein interactionComputational biologyBiochemistryBiophysicsChromatographyBiologyUbiquitin and proteasome pathwaysAdvanced Proteomics Techniques and ApplicationsMass Spectrometry Techniques and Applications