Abstract
Studies on the specificity and mechanism of pepsin action have involved the use of several types of synthetic substrates. In the early work, substrates related to Z-Glu-Tyr (1) or Ac-Phe-Tyr (2) were largely employed; for a review, see (3). In particular, much use had been made of acyl dipeptides of the type A-X-Y, where X and Y are aromatic L-amino acid residues forming the sensitive peptide bond. Because the pKa of the carboxyl group falls in the pH range of pepsin activity, the pH dependence of the kinetic parameters is a function of the ionization of prototrophic groups in both the enzyme and the substrate. The methyl or ethyl esters, or amides, of acyl dipeptides have also been used extensively, but because of their limited solubility in aqueous solution, variable amounts of organic solvents had to be added. Such solvents, even in relatively low concentration, markedly inhibit pepsin action (4,5). To obviate this difficulty, another type of pepsin substrate was introduced, in which a cationic group (the imidazolium group of a His residue, the α-ammonium group, a pyridinium group, or a morpholinium group) is present. By means of such substrates, in particular of the general structure Z-His-X-Y-OMe, the primary specificity of pepsin was defined as a preference for hydrophobic L-amino acid residues in both the X- and Y- positions (10); substitution of either Phe residue of Z-His-Phe-Phe-OMe by its D-enatiomer renders the X-Y bond resistant to pepsin action (6). The favorable effect of an aromatic and planar substituent at the β-carbon of the X and Y residues was emphasized by the finding that when X is β-cyclohexyl-L-alanyl, the value of kcat is much lower than that found for the corresponding substrate in which X or Y = Phe, and is similar to that for substrates in which the X- or Y-position is occupied by an aliphatic amino acid residue larger than Ala (Nva, Nle, Leu, Met). Apparently, the side chains of these amino acids can interact with a portion of the enzymic region that binds planar aromatic groups. It was also shown that the replacement of Phe in the X-position by Val or Ile rendered the X-Y bond resistant to pepsin action than when X = Gly, indicating that when the X-position is occupied by a residue that is branched at the β-carbon, one of the catalytic groups of pepsin may be prevented from attacking the carbonyl group of the sensitive bond. Moreover, the importance of the β-methylene group as a structural element of the X-Y unit was underlined by the finding that replacement of either Phe residue of Z-His-Phe-Phe-OMe by a L-Phenylglycyl residue also rendered the X-Y bond resistant to pepsin action (11).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Fruton, J. S., and Bergmann, M. (1939) J. Biol. Chem. 127, 627–641
Baker, L. E. (1951) J. Biol. Chem. 193, 809–819
Clement, G. E. (1973) Progr. Bioorg. Chem. 2, 177–238
Tang, J. (1965) J. Biol. Chem. 240, 3810–3815
Zeffren, E., and Kaiser, E. T. (1967) J. Amer. Chem. Soc. 89, 4204–4208
Inouye, K., and Fruton, J. S. (1967) Biochem. 6, 1765–1777
Hollands, T. R., Voynick, I. M., and Fruton, J. S. (1969) Biochem. 8, 575–585
Sachdev, G. P., and Fruton, J. S. (1969) Biochem. 8, 4231–4238
Tikhodeeva, A. G., Rumsh, L. D., and Antonov, V. K. (1975) Bioorg. Khimia 1, 993–994
Trout, G. E., and Fruton, J. S. (1969) Biochem. 8, 4183–4190
Voynick, I. M., and Fruton, J. S., unpublished experiments
Rumsh, L. D., Tikhodeeva, A. G., and Antonov, V. K. (1974) Biokhimiya 39, 899–902
Fruton, J. S. (1976) Adv. Enzymol. 44, 1–36
Sampath-Kumar, P. S., and Fruton, J. S. (1974) Proc. Natl. Acad. Sci. U.S.A. 71, 1070–1072
Schechter, I., and Berger, A. (1967) Biochem. Biophys. Res. Commun. 27, 157–162
Lowbridge, J., and Fruton, J. S. (1974) J. Biol. Chem. 249, 6754–6761
Sachdev, G. P., and Fruton, J. S. (1970) Biochem. 9, 4465–4470
Sachdev, G. P., Johnston, M. A., and Fruton, J. S. (1972) Biochem. 11, 1080–1086
Sachdev, G. P., Brownstein, A. D., and Fruton, J. S. (1973) J. Biol. Chem. 248, 6292–6299
Sachdev, G. P., Brownstein, A. D., and Fruton, J. S. (1975) J. Biol. Chem. 250, 501–507
Humphreys, R. E., and Fruton, J. S. (1968) Proc. Natl. Acad Sci. U.S.A. 59, 519–525
Raju, E. V., Humphreys, R. E., and Fruton, J. S. (1972) Biochem. 11, 3533–3536
Wang, J. L., and Edelman, G. M. (1971) J. Biol. Chem. 246, 1185–1191
Sachdev, G. P., and Fruton, J. S. (1975) Proc. Natl. Acad. Sci. U.S.A. 72, 3424–3427
Press, E. M., Porter, R. R., and Cebra, J. (1960) Biochem. J. 74, 501–514
Woessner, J. F., and Shamberger, R. J. (1971) J. Biol. Chem. 246, 1951–1960
Voynick, I. M., and Fruton, J. S. (1971) Proc. Natl. Acad. Sci. U.S.A. 68, 257–259
Raymond, M. N., Garnier, J., Bricas, E., Cilianu, S., Blasnic, M. Chaix, A., and Lefrancier, P. (1972) Biochimie 54, 145–154
Fruton, J. S. (1975) in Proteases and Biological Control (Reich, E., Rifkin, D. B., and Shaw, E., Eds) pp. 33–50, Cold Spring Harbor Laboratory, New York
Inouye, K., and Fruton, J. S. (1967) J. Amer. Chem. Soc. 89, 187–188
May, S. W., and Kaiser, E. T. (1969) J. Amer. Chem. Soc. 91, 6491–6495
May, S. W., and Kaiser, E. T. (1972) Biochem. 11, 592–600
Hunkapiller, M. W., and Richards, J. H. (1972) Biochem. 11, 2829–2839
Takahashi, M., Want, T. T., and Hofmann, T. (1974) Biochem. Biophys. Res. Commun. 57, 39–46
Bender, M., and Kezdy, F. (1965) Ann. Rev. Biochem. 34, 49–76
Thompson, R. C., and Blout, E. R. (1973) Biochem. 12, 57–65
Jencks, W. P. (1975) Adv. Enzymol. 43, 219–410
Burgen, A. S. V., Roberts, G. C. R., and Feeney, J. (1975) Nature 253, 753
Mattis, J. A., and Fruton, J. S. (1976) Biochem. 15, 2191–2194
Sepulveda, P., Marciniszyn, J., Liu, D., and Tang, J. (1975) J. Biol. Chem. 250, 5082–5088
Bender, M. L., Chow, Y., and Chloupek, F. (1958) J. Amer. Chem. Soc. 80, 5380–5387
Kirby, A. J., McDonald, R. S., and Smith, C. R. (1974) J. Chem. Soc., Perkin Trans. 2 1974, 1495–1504
Wolfenden, R. (1976) Ann. Rev. Biophys. Bioeng. 5, 271–306
Marciniszyn, J. P., Hartsuck, J. A., and Tang, J. (1975) Fed. Proc. 34, 484
Rich, D. H., Sun, E., and Singh, J. (1976) Abstr. Amer. Chem. Soc. Divn. Biol. Chem., 172nd Meeting, San Francisco, Cal., No. 42
Cornish-Bowden, A. J., Greenwell, P., and Knowles, J. R. (1969) Biochem. J. 113, 369–375
Takahashi, M., and Hofmann, T. (1975) Biochem. J. 147, 549–563
Newmark, A. K., and Knowles, J. R. (1975) J. Amer. Chem. Soc. 97, 3557–3559
Nakagawa, Y., King Sun, L. H., and Kaiser, E. T. (1976) J. Amer. Chem. Soc. 98, 1616–1617
Zeffren, E., and Kaiser, E. T. (1968) Arch. Biochem. Biophys. 126, 965–967
Silver, M. S., and Stoddard, M. (1972) Biochem. 11, 191–200
Silver, M. S., Stoddard, M., and Kelleher, M. H. (1976) J. Amer. Chem. Soc. 98, (in press)
Richman, P. G., and Fruton, J. S. (1976) Proc. Natl. Acad. Sci. U.S.A. 73, (in press)
Determann, H., Heuer, J., and Jaworek, D. (1965) Ann. Chem. 690, 189–196
Kozlov, L. V., Ginodman, L. M., Orekhovich, V. N., and Valueva, T. A. (1966) Biokhymiya 31, 315–321
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1977 Springer Science+Business Media New York
About this chapter
Cite this chapter
Fruton, J.S. (1977). Specificity and Mechanism of Pepsin Action on Synthetic Substrates. In: Tang, J. (eds) Acid Proteases: Structure, Function, and Biology. Advances in Experimental Medicine and Biology, vol 95. Springer, New York, NY. https://doi.org/10.1007/978-1-4757-0719-9_8
Download citation
DOI: https://doi.org/10.1007/978-1-4757-0719-9_8
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4757-0721-2
Online ISBN: 978-1-4757-0719-9
eBook Packages: Springer Book Archive