A strategy for dissecting the architectures of native macromolecular assemblies

Yi Shi, Riccardo Pellarin, Peter C. Fridy, Javier Fernandez-Martinez, Mary K. Thompson, Yinyin Li, Qing Jun Wang, Andrej Sali, Michael P. Rout, Brian T. Chait

Research output: Contribution to journalArticlepeer-review

81 Scopus citations

Abstract

It remains particularly problematic to define the structures of native macromolecular assemblies, which are often of low abundance. Here we present a strategy for isolating complexes at endogenous levels from GFP-tagged transgenic cell lines. Using cross-linking mass spectrometry, we extracted distance restraints that allowed us to model the complexes' molecular architectures.

Original languageEnglish
Pages (from-to)1135-1138
Number of pages4
JournalNature Methods
Volume12
Issue number12
DOIs
StatePublished - Dec 1 2015

Bibliographical note

Funding Information:
We are grateful to A.N. Krutchinsky (The Rockefeller University, New York, New York, USA) for providing the yeast GFP-CDC16 strain; S. Obado for technical assistance; E. Jacobs, J. Lacava, S.J. Kim and B. Webb for discussions and assistance; and Z. Yue (Icahn School of Medicine at Mount Sinai, New York, New York, USA) for sharing Becn1-EGFP/+ mice. Y.S. acknowledges M. Chen (R.G. Roeder lab, The Rockefeller University, New York, New York, USA) for generating a reagent that was not used for this study. This work was funded by the US National Institutes of Health (grants P41 GM103314 (to B.T.C.), R01 GM083960 (to A.S.) and P41 GM109824 (to A.S., M.P.R. and B.T.C.)) and the Ellison Medical Foundation (Q.J.W.).

Publisher Copyright:
© 2015 Nature America, Inc.

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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