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

Franz Herzog, Abdullah Kahraman, Daniel Boehringer, Raymond Mak, Andreas Bracher, Thomas Walzthoeni, Alexander Leitner, Martin Beck, Franz-Ulrich Hartl, Nenad Ban, Lars Malmström, Ruedi Aebersold

Department of Biology, Institute of Molecular Systems Biology, Eidgenössische Technische Hochschule Zürich, Wolfgang-Pauli Strasse 16, 8093 Zurich, Switzerland.

Science (New York, N.Y.) 2012 Sep 14

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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.

Citation

Franz Herzog, Abdullah Kahraman, Daniel Boehringer, Raymond Mak, Andreas Bracher, Thomas Walzthoeni, Alexander Leitner, Martin Beck, Franz-Ulrich Hartl, Nenad Ban, Lars Malmström, Ruedi Aebersold. Structural probing of a protein phosphatase 2A network by chemical cross-linking and mass spectrometry. Science (New York, N.Y.). 2012 Sep 14;337(6100):1348-52