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Proteolytic Enzymes for Peptide Production

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Enzymes of Molecular Biology

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 16))

Abstract

There are three main reasons why a protein chemist might wish to cleave a protein of interest into peptide fragments. The first reason is to generate, by extensive proteolysis, a large number of relatively small (5–20 residues) peptides either for peptide mapping (see vol. 1, Chapter 5) or for purification and subsequent manual sequence determination by the dansyl-Edman method (see vol. 1, Chapter 24). The second reason is to generate relatively large peptides (50–150 residues) by limited proteolysis for automated sequence analysis, such as with the gas-phase sequencer. The third reason is to prepare, again by limited proteolysis, specific fragments for studies relating structure to function, In each case, the specificity of the enzyme used to generate the peptides is a prime consideration, since the aim is to provide high yields of discrete fragments. It can be appreciated that significantly <100% cleavage at some or all of the cleavage sites on the protein being digested will generate a far more complex mixture of a larger number of polypeptides, each in relatively low yield. It is for this reason that enzymes of high specificity, such as trypsin, which cleaves at the C-terminal side of arginine and lysine residues, are mainly used for peptide production.

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Authors and Affiliations

  1. Division of Biological Sciences, The University of Hertfordshire, Hatfield, UK

    Patricia J. Sweeney & John M. Walker

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  1. Patricia J. Sweeney
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  2. John M. Walker
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  1. Advanced Technologies (Cambridge) Limited, Cambridge, UK

    Michael M. Burrell

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© 1993 Humana Press Inc.

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Sweeney, P.J., Walker, J.M. (1993). Proteolytic Enzymes for Peptide Production. In: Burrell, M.M. (eds) Enzymes of Molecular Biology. Methods in Molecular Biology™, vol 16. Humana Press. https://doi.org/10.1385/0-89603-234-5:277

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  • DOI: https://doi.org/10.1385/0-89603-234-5:277

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-322-1

  • Online ISBN: 978-1-59259-503-7

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