Targeted cross-linking-mass spectrometry determines vicinal interactomes within heterogeneous RNP complexes.

ABSTRACT: Proteomic and RNomic approaches have identified many components of different ribonucleoprotein particles (RNPs), yet still little is known about the organization and protein proximities within these heterogeneous and highly dynamic complexes. Here we describe a targeted cross-linking approach, which combines cross-linking from a known anchor site with affinity purification and mass spectrometry (MS) to identify the changing vicinity interactomes along RNP maturation pathways. Our method confines the reaction radius of a heterobifunctional cross-linker to a specific interaction surface, increasing the probability to capture low abundance conformations and transient vicinal interactors too infrequent for identification by traditional cross-linking-MS approaches, and determine protein proximi

SUBMITTER: Trahan C 

PROVIDER: S-EPMC4756821 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature