Quantitative NMR analysis of the protein G B1 domain in Xenopus laevis egg extracts and intact oocytes

Philipp Selenko, Zach Serber, Bedrick Gadea, Joan Ruderman, Gerhard Wagner

Research output: Contribution to journalArticlepeer-review

200 Scopus citations

Abstract

We introduce a eukaryotic cellular system, the Xenopus laevis oocyce, for in-cell NMR analyses of biomolecules at high resolution and delineate the experimental reference conditions for successful implementations of in vivo NMR measurements in this cell type. This approach enables quantitative NMR experiments at defined intracellular concentrations of exogenous proteins, which is exemplified by the description of in-cell NMR properties of the protein G B1 domain (GB1). Additional experiments in Xenopus egg extracts and artificially crowded in vitro solutions suggest that for this biologically inert protein domain, intracellular viscosity and macromolecular crowding dictate its in vivo behavior. These contributions appear particularly pronounced for protein regions with high degrees of internal mobility in the pure state. We also evaluate the experimental limitations of this method and discuss potential applications toward the in situ structural characterization of eukaryotic cellular activities.

Original languageEnglish (US)
Pages (from-to)11904-11909
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number32
DOIs
StatePublished - Aug 8 2006
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • General

Keywords

  • High-resolution
  • In-cell
  • Liquid-state

Fingerprint

Dive into the research topics of 'Quantitative NMR analysis of the protein G B1 domain in Xenopus laevis egg extracts and intact oocytes'. Together they form a unique fingerprint.

Cite this