RRC ID 21532
著者 Ito Y, Mikawa T, Smith BO.
タイトル In-cell NMR of intrinsically disordered proteins in prokaryotic cells.
ジャーナル Methods Mol Biol
Abstract In-cell NMR, i.e., the acquisition of heteronuclear multidimensional NMR of biomacromolecules inside living cells, is, to our knowledge, the only method for investigating the three-dimensional structure and dynamics of proteins at atomic detail in the intracellular environment. Since the inception of the method, intrinsically disordered proteins have been regarded as particular targets for in-cell NMR, due to their expected sensitivity to the molecular crowding in the intracellular environment. While both prokaryotic and eukaryotic cells can be used as host cells for in-cell NMR, prokaryotic in-cell NMR, particularly employing commonly used protein overexpression systems in Escherichia coli cells, is the most accessible approach. In this chapter we describe general procedures for obtaining in-cell NMR spectra in E. coli cells.
巻・号 895
ページ 19-31
公開日 2012-1-1
DOI 10.1007/978-1-61779-927-3_2
PMID 22760309
MeSH Culture Techniques Escherichia coli / chemistry* Escherichia coli / metabolism Isotope Labeling Nuclear Magnetic Resonance, Biomolecular* Prokaryotic Cells / metabolism Protein Conformation Recombinant Proteins / biosynthesis Recombinant Proteins / chemistry
リソース情報
遺伝子材料 Thermus thermophilus expression plasmid TEx14F08 (THR005728) TEx02C08 (THR000856) TEx16E07 (THR006503).