Literature DB >> 22039762

Specific labeling of threonine methyl groups for NMR studies of protein-nucleic acid complexes.

Kaustubh Sinha1, Linda Jen-Jacobson, Gordon S Rule.   

Abstract

Specific (13)C labeling of Thr methyl groups has been accomplished via the growth of a standard laboratory strain of Escherichia coli on [2-(13)C]glycerol in the presence of deuterated isoketovalerate, Ile, and Ala. Diversion of the label from the Thr biosynthetic pathway is suppressed by including Lys, Met, and Ile in the growth medium. This method complements the repertoire of methyl labeling schemes for NMR structural and dynamic studies of proteins and is particularly useful for the study of nucleic acid binding proteins because of the high propensity of Thr residues at protein-DNA and -RNA interfaces.

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Year:  2011        PMID: 22039762      PMCID: PMC3222749          DOI: 10.1021/bi201496d

Source DB:  PubMed          Journal:  Biochemistry        ISSN: 0006-2960            Impact factor:   3.162


  21 in total

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9.  Cross-correlated relaxation enhanced 1H[bond]13C NMR spectroscopy of methyl groups in very high molecular weight proteins and protein complexes.

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4.  Bacterial production of site specific 13C labeled phenylalanine and methodology for high level incorporation into bacterially expressed recombinant proteins.

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Review 6.  The quiet renaissance of protein nuclear magnetic resonance.

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Review 8.  Practical aspects of NMR signal assignment in larger and challenging proteins.

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10.  An economical method for production of (2)H, (13)CH3-threonine for solution NMR studies of large protein complexes: application to the 670 kDa proteasome.

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  10 in total

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