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Chemical Cleavage of Proteins at Tryptophanyl-X Peptide Bonds

  • Bryan John Smith
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Tryptophan is represented in the genetic code by a single codon and has proven useful in cloning exercises in providing an unambiguous oligonucleotide sequence as part of a probe or primer. It is also one of the less abundant amino acids found in polypeptides, and cleavage of bonds involving tryptophan generates large peptides. This may be convenient for generation of internal sequence information (the usual purpose to which this technique is put), but is less useful for identification of proteins by mass mapping, where a larger number of smaller peptides makes for more successful database searching.

Keywords

Fume Hood Leader Peptide Cyanogen Bromide Usual Purpose Large Peptide 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Kilic, F. and Ball, E. H. (1991). Partial cleavage mapping of the cytoskeletal protein vinculin. Antibody and talin binding sites. J. Biol. Chem. 266, 8734–8740.PubMedGoogle Scholar
  2. 2.
    Litchfield, D. W., Lozeman, F. J., Cicirelli, M. F., Harrylock, M., Ericsson, L. H., Piening, C. J., and Krebs. E. G. (1991) Phosphorylation of the beta subunit of casein kinase II in human A431 cells. Identification of the autophosphorylation site and a site phosphorylated by p34cdc2. J. Biol. Chem. 266, 20,380–20,389.PubMedGoogle Scholar
  3. 3.
    Tseng, A., Buchta, R., Goodman, A. E., Loughman, M., Cairns D., Seilhammer, J., et al. (1991) A strategy for obtaining active mammalian enzyme from a fusion protein expressed in bacteria using phospholipase A2 as a model. Pro. Express. and Purificat. 2, 127–135.CrossRefGoogle Scholar
  4. 4.
    Huang, H. V., Bond, M. W., Hunkapillar, M. W., and Hood, L. E. (1983) Cleavage at tryptophanyl residues with dimethyl sulfoxide-hydrochloric acid and cyanogen bromide. Meth. Enzymol. 91, 318–324.PubMedCrossRefGoogle Scholar
  5. 5.
    Rosa, J. C., de Oliveira, P. S. L., Garrat, R., Beltramini, L., Roque-Barreira, M.-C. and Greene, L. J. (1999) KM+, a mannose-binding lectin from Artocarpus integrifolia: amino acid sequence, predicted tertiary structure, carbohydrate recognition, and analysis of the beta-prism fold. Prot. Sci. 8, 13–24.CrossRefGoogle Scholar
  6. 6.
    Beavis, R. C. and Chait. B. T. (1990). Rapid, sensitive analysis of protein mixtures by mass spectrometry. Proc. Natl. Acad. Sci. USA 87, 6873–6877.PubMedCrossRefGoogle Scholar
  7. 7.
    Morrison, J. R., Fidge, N. H., and Grego, B. (1990) Studies on the formation, separation, and characterisation of cyanogen bromide fragments of human A1 apolipoprotein. Analyt. Biochem. 186, 145–152.PubMedCrossRefGoogle Scholar
  8. 8.
    Ozols, J. and Gerard, C. (1977). Covalent structure of the membranous segment of horse cytochrome b5. Chemical cleavage of the native hemprotein. J. Biol. Chem. 252, 8549–8553.PubMedGoogle Scholar
  9. 9.
    Savige, W. E. and Fontana, A. (1977) Cleavage of the tryptophanyl peptide bond by dim-ethyl sulfoxide-hydrobromic acid. Meth. Enzymol. 47, 459–469.PubMedCrossRefGoogle Scholar
  10. 10.
    Fontana, A. (1972) Modification of tryptophan with BNPS skatole (2-(2-nitro-phenylsulfenyl)-3-methyl-3′-bromoindolenine). Meth. Enzymol. 25, 419–423.CrossRefGoogle Scholar
  11. 11.
    Crimmins, D. L., McCourt, D. W., Thoma, R. S., Scott, M. G., Macke, K. and Schwartz, B. D. (1990) In situ cleavage of proteins immobilised to glass-fibre and polyvinylidene difluoride membranes: cleavage at tryptophan residues with 2-(2′-nitropheylsulfenyl)-3-methyl-3′-bromoindolenine to obtain internal amino acid sequence. Analyt. Biochem. 187, 27–38.PubMedCrossRefGoogle Scholar
  12. 12.
    Ramachandran, L. K. and Witkop, B. (1976).-Bromosuccinimide cleavage of peptides. Meth. Enzymol. 11, 283–299.CrossRefGoogle Scholar
  13. 13.
    Lischwe, M. A. and Sung, M. T. (1977). Use of-chlorosuccinimide/urea for the selective cleavage of tryptophanyl peptide bonds in proteins. J. Biol. Chem. 252, 4976–4980.PubMedGoogle Scholar
  14. 14.
    Lischwe, M. A. and Ochs, D. (1982). A new method for partial peptide mapping using chlorosuccinimide/urea and peptide silver staining in sodium dodecyl sulphate-polyacry-lamide gels. Analyt. Biochem. 127, 453–457.PubMedCrossRefGoogle Scholar
  15. 15.
    Fontana, A., Dalzoppo, D., Grandi, C., and Zambonin, M. (1983) Cleavage at tryptophan with iodosobenzoic acid. Meth. Enzymol. 91, 311–318.PubMedCrossRefGoogle Scholar
  16. 16.
    Vestling, M. M., Kelly, M. A., and Fenselau, C. (1994) Optimization by mass spectrom-etry of a tryptophan-specific protein cleavage reaction. Rapid Commun. Mass Spectr. 8, 786–790.CrossRefGoogle Scholar
  17. 17.
    Rahali, V. and Gueguen, J. (1999) Chemical cleavage of bovine (β-lactoglobulin by BNPS-skatole for preparative purposes: comparative study of hydrolytic procedures and peptide characterization. J. Prot. Chem. 18, 1–12.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Bryan John Smith
    • 1
  1. 1.Celltech, R&DSloughUK

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