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Structural probing of a protein phosphatase 2A network by chemical cross-linking and mass spectrometry

Science. 2012 Sep 14;337(6100):1348-52. doi: 10.1126/science.1221483.

Abstract

The identification of proximate amino acids by chemical cross-linking and mass spectrometry (XL-MS) facilitates the structural analysis of homogeneous protein complexes. We gained distance restraints on a modular interaction network of protein complexes affinity-purified from human cells by applying an adapted XL-MS protocol. Systematic analysis of human protein phosphatase 2A (PP2A) complexes identified 176 interprotein and 570 intraprotein cross-links that link specific trimeric PP2A complexes to a multitude of adaptor proteins that control their cellular functions. Spatial restraints guided molecular modeling of the binding interface between immunoglobulin binding protein 1 (IGBP1) and PP2A and revealed the topology of TCP1 ring complex (TRiC) chaperonin interacting with the PP2A regulatory subunit 2ABG. This study establishes XL-MS as an integral part of hybrid structural biology approaches for the analysis of endogenous protein complexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chaperonins / chemistry
  • Cross-Linking Reagents / chemistry
  • Crystallography, X-Ray
  • Humans
  • Mass Spectrometry / methods*
  • Metabolic Networks and Pathways*
  • Protein Conformation
  • Protein Interaction Mapping / methods*
  • Protein Phosphatase 2 / chemistry*

Substances

  • Cross-Linking Reagents
  • Protein Phosphatase 2
  • Chaperonins