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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| Published in | Journal of biomolecular NMR Vol. 43; no. 4; pp. 229 - 238 |
|---|---|
| Main Authors | , , |
| Format | Journal Article |
| Language | English |
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Dordrecht
Dordrecht : Springer Netherlands
2009
Springer Netherlands |
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| Abstract | 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. |
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| AbstractList | 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
1
H–
13
C 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
13
CH
3
sites on a deuterated background. 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. |
| Author | Guo, Chenyun Sheppard, Devon Tugarinov, Vitali |
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| Cites_doi | 10.1021/ja0205636 10.1006/jmbi.1996.0699 10.1021/ja030345s 10.1002/mrc.1256 10.1038/nature05512 10.1021/bi992519h 10.1007/BF00197809 10.1023/B:JNMR.0000013824.93994.1f 10.1021/ja030153x 10.1021/ja00510a007 10.1146/annurev.biochem.73.011303.074004 10.1023/A:1008393201236 10.1007/s10858-008-9285-8 10.1038/nmeth1080 10.1021/ja808202q 10.1021/ja053539b 10.1006/jmbi.1996.0603 10.1073/pnas.0703347104 10.1021/ja052517m 10.1146/annurev.biophys.27.1.357 10.1021/ja0761784 10.1006/jmra.1994.1213 10.1038/nmeth938 10.1021/bi9624806 10.1021/ja00032a026 10.1007/BF00404272 10.1002/prot.340040204 10.1021/jp0348124 |
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| Keywords | Methyl assignments Methyl-TROSY Isotope labeling Large proteins |
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| Snippet | A set of sensitive methyl-detected 'out-and-back' NMR experiments for simultaneous assignments of Alaβ and Ileγ2 methyl positions in large proteins is... A set of sensitive methyl-detected ‘out-and-back’ NMR experiments for simultaneous assignments of Alaβ and Ileγ2 methyl positions in large proteins is... |
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| SubjectTerms | Biochemistry Biological and Medical Physics Biophysics Physics Physics and Astronomy Spectroscopy/Spectrometry |
| Title | Methyl-detected 'out-and-back' NMR experiments for simultaneous assignments of Alaβ and Ileγ2 methyl groups in large proteins |
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