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TROSY NMR Spectroscopy of Large Soluble Proteins

  • Yingqi Xu
  • Stephen MatthewsEmail author
Chapter
Part of the Topics in Current Chemistry book series (TOPCURRCHEM, volume 335)

Abstract

Solution nuclear magnetic resonance spectroscopy is usually only used to study proteins with molecular weight not exceeding about 50 kDa. This size limit has been lifted significantly in recent years, thanks to the development of labelling methods and the application of transverse-relaxation optimized spectroscopy (TROSY). In particular, methyl-specific labelling and methyl-TROSY have been shown to be effective for supramolecular systems as large as about 1 MDa. In this chapter we review the available methods for labelling different kinds of methyl groups and the assignment strategies in very large protein systems. Several proteins are selected as examples to show how NMR helps to reveal the details of structure, interaction and dynamics of these proteins.

Keywords

Large proteins Methyl labelling Mutagenesis NOESY TROSY 

Abbreviations

ATCase

Aspartate transcarbamoylase

CSA

Chemical shift anisotropy

HMQC

Heteronuclear multiple-quantum correlation

HSQC

Heteronuclear single-quantum correlation

NMR

Nuclear magnetic resonance

NOE

Nuclear Overhauser effect

PCS

Pseudocontact shift

PRE

Paramagnetic relaxation enhancement

RDC

Residual dipolar coupling

TROSY

Transverse relaxation optimized spectroscopy

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Copyright information

© Springer-Verlag Berlin Heidelberg 2011

Authors and Affiliations

  1. 1.Division of Molecular BiosciencesImperial College LondonLondonUK

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