Methyl-detected 'out-and-back' NMR experiments for simultaneous assignments of Alaβ and Ileγ2 methyl groups in large proteins

A set of sensitive methyl-detected 'out-and-back' NMR experiments for simultaneous assignments of Alaβ and Ileγ2 methyl positions in large proteins is described. The developed methodology is applied to an 82-kDa enzyme Malate Synthase G. Complete alanine β and isoleucine γ2 ¹H-¹³C methyl c...

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Bibliographic Details
Published inJournal of biomolecular NMR Vol. 43; no. 4; pp. 229 - 238
Main Authors Sheppard, Devon, Guo, Chenyun, Tugarinov, Vitali
Format Journal Article
LanguageEnglish
Published Dordrecht Dordrecht : Springer Netherlands 2009
Springer Netherlands
Subjects
Biochemistry
Biological and Medical Physics
Biophysics
Physics
Physics and Astronomy
Spectroscopy/Spectrometry
Methyl assignments
Methyl-TROSY
Isotope labeling
Large proteins
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Summary:A set of sensitive methyl-detected 'out-and-back' NMR experiments for simultaneous assignments of Alaβ and Ileγ2 methyl positions in large proteins is described. The developed methodology is applied to an 82-kDa enzyme Malate Synthase G. Complete alanine β and isoleucine γ2 ¹H-¹³C methyl chemical shift assignments could be obtained from the set of new methyl-detected 'out-and-back' 3D experiments. The described methodology for methyl assignments should be applicable to protein molecules within approximately 100-kDa molecular weight range irrespective of the labeling strategy chosen to produce selectively protonated Alaβ and Ileγ2 ¹³CH₃ sites on a deuterated background.
Bibliography:http://dx.doi.org/10.1007/s10858-009-9305-3
ISSN:0925-2738
1573-5001
DOI:10.1007/s10858-009-9305-3