Chemical Modification of Lysine by Reductive Methylation

A Probe for Residues Involved in DNA Binding
  • Ian A. Taylor
  • Michelle Webb
Part of the Methods in Molecular Biology book series (MIMB, volume 148)


The basic side chains of lysine residues often play essential roles in DNA-protein recognition. They are able to contribute to the overall affinity of an interaction through nonspecific charge-charge interactions with the phosphate backbone and contribute substantially to the specificity of the interaction by forming direct hydrogen bonds with functional groups on the edges of the bases. This dual role and their almost ubiquitous presence in the interface of DNA-protein complexes make them very attractive targets for chemical modification experiments.


Lysine Residue Liquid Scintillant Lysine Side Chain Sodium Cyanoborohydride Incorporated Radioactivity 
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  1. 1.
    Lundblad, R. L. and Noyes, C. M. (1984) Chemical Reagents for Protein Modification. CRC, Boca Raton, FL.Google Scholar
  2. 2.
    Hunter, M. J. and Ludwig, M. L. (1962) J. Am. Chem. Soc. 84, 3491–3497.CrossRefGoogle Scholar
  3. 3.
    Means, G. E. and Feeney, R. E. (1968) Reductive alkylation of amino groups in proteins. Biochemistry 7, 2192–2201.PubMedCrossRefGoogle Scholar
  4. 4.
    Means, G. E. and Feeney, R. E. (1995) Reductive alkylation of proteins. Anal. Biochem. 224, 1–16.PubMedCrossRefGoogle Scholar
  5. 5.
    Jentoft, N. and Dearborn, D. G. (1983) Protein labelling by reductive alkylation. Methods Enzymol. 91, 570–579.PubMedCrossRefGoogle Scholar
  6. 6.
    Jentoft, N. and Dearborn, D. G. (1979) Labelling of proteins by reductive methylation using sodium cyanoborohydride. J. Biol. Chem. 254, 4359–4365.PubMedGoogle Scholar
  7. 7.
    Zhang, M., Thulin, E., and Vogel, H. J. (1994) Reductive methylation and pKa determination of the lysine side chains in calbindin D9k. J. Protein Chem. 13, 527–535.PubMedCrossRefGoogle Scholar
  8. 8.
    Lambert, S. F. and Thomas, J. O. (1986) Lysine-containing DNA-binding regions on the surface of the histone octamer in the nucleosome core particle. Eur. J. Biochem. 160, 191–201.PubMedCrossRefGoogle Scholar
  9. 9.
    Thomas, J. O. and Wilson, C. M. (1986) Selective radiolabelling and identification of a strong nucleosome binding site on the globular domain of histone H5. EMBOJ. 5, 3531–3537.Google Scholar
  10. 10.
    Taylor, I. A., Webb, M., and Kneale, G. G. (1996) Surface labeling of the type I methyltransferase M.EcoR124I reveals lysine residues critical for DNA binding. J. Mol. Biol. 258, 62–73.PubMedCrossRefGoogle Scholar
  11. 11.
    Poncz, L. and Dearborn, D. G. (1983) The resistance to tryptic hydrolysis of peptide bonds adjacent to N-epsilon, N-dimethyllysyl residues. J. Biol. Chem. 258, 1844–1850.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2001

Authors and Affiliations

  • Ian A. Taylor
    • 1
  • Michelle Webb
    • 1
  1. 1.Department of ChemistryUniversity of SheffieldSheffieldUK

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