Abstract
The β-turn formed by the amino acid residues 20–23 of the B-chain of insulin has been implicated as an important structural feature of the molecule. In other biologically active peptides, stabilization of β-turns has resulted in increases in activity. We have synthesized three insulin analogues containing modifications which would be expected to increase the stability of the β-turn. In two analogues, we have substituted α-aminoisobutyric acid (Aib) for the Glu residue normally present in position B21 or for the Arg residue normally present in position B22; in a third compound, we have replaced the Glu residue with its D-isomer. Biological evaluation of these compounds showed that [B21 Aib]insulin displays a potencyca. one-fourth that of natural insulin, while [B22 Aib]insulin is less than 10% as potent. In contrast, [B21 D-Glu]insulin is equipotent with natural insulin. We conclude that the β-turn region of the insulin molecule normally possesses considerable flexibility, which may be necessary for it to assume a conformation commensurate with high biological activity. If this is the case, [B21 D-Glu]insulin may exhibit a stabilized geometry similar to that of natural insulin when bound to the insulin receptor.
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References
Anderson, G. W., Zimmerman, J. E., and Callahan, F. M. (1964).J. Am. Chem. Soc. 86, 1839–1842.
Barany, G., and Merrifield, R. B. (1980). InThe Peptides, Gross, E., and Meienhofer, J. (eds.), Academic Press, New York, Vol. 2, pp. 3–284.
Blundell, T., Dodson, G. G., Dodson, E., Hodgkin, D. C., and Vigayan, M. (1971).Rec. Prog. Horm. Res. 27, 1–40.
Blundell, T., Dodson, G. G., Hodgkin, D. C., and Mercola, D. (1972).Adv. Protein Chem. 26, 279–402.
Casaretto, M., Spoden, M., Diconescu, C., Gattner, H.-G., Zahn, H., Brandenburg, D., and Wollmer, A. (1987).Biol. Chem. Hoppe-Seyler 368, 709–716.
Chance, R. E., Hoffman, J. A., Kroef, E. R., Johnson, M. G., Schirmer, E. W., Bromer, W. W., Ross, M. J., and Wetzel, R. (1981).Proc. Am. Pept. Symp. 7, 721–728.
Chou, P. Y., and Fasman, G. D. (1978).Adv. Enzymol. 47, 45–148.
Cutfield, J. F., Cutfield, S. M., Dodson, E. J., Dodson, G. G., Reynolds, D. D., and Vallely, D. (1981). InStructural Studies on Molecules of Biological Interest, Dodson, G., Gluscker, J. P., and Sayre, D. (eds.), Clarendon Press, Oxford, pp. 527–546.
Freidinger, R. M. (1981).Proc. Am. Pept. Symp. 7, 673–683.
Hagenmaier, H., and Frank, H. (1972).Hoppe-Seyler's Z. Physiol. Chem. 353, 1973–1976.
Isogai, Y., Nemethy, G., and Scheraga, H. A. (1977).Proc. Natl. Acad. Sci. USA 74, 414–418.
Jorgenson, E. C., Rapaka, S.-P., Windridge, G. C., and Lee, T. C. (1971).J. Med. Chem. 14, 904–906.
Joshi, S., Burke, G. T., and Katsoyannis, P. G. (1985).Biochemistry 24, 4208–4214.
Joshi, S., Burke, G. T., and Katsoyannis, P. G. (1990).J. Protein Chemistry 9, 235–246.
Kaiser, E., Colescott, R. L., Bossinger, C. D., and Cook, P. I. (1970).Anal. Biochem. 34, 595–598.
Katsoyannis, P. G., Tometsko, A., Zalut, C., Johnson, S., and Trakatellis, A. C. (1967a).Biochemistry 6, 2635–2641.
Katsoyannis, P. G., Trakatellis, A. C., Johnson, S., Zalut, C., and Schwartz, G. P. (1967b).Biochemistry 6, 2642–2655.
Katsoyannis, P. G., Trakatellis, A. C., Zalut, C., Johnson, S., Tometsko, A., Schwartz, G. P., and Ginos, J. (1967c).Biochemistry 6, 2656–2668.
Katsoyannis, P. G., Ginos, J., Cosmatos, A., and Schwartz, G. P. (1975).J. Chem. Soc. Perkin Trans. 1, 464–469.
Katsoyannis, P. G., and Schwartz, G. P. (1977).Methods Enzymol. 47, 501–578.
Kitagawa, K., Ogawa, H., Burke, G. T., Chanley, J. D., and Katsoyannis, P. G. (1984).Biochemistry 23, 1405–1413.
Kobayashi, M., Ohgaku, S., Iwasaki, M., Maegawa, H., Shigeta, Y., and Inouye, K. (1982).Biochem. Biophys. Res. Commun. 107, 329–336.
Krstenansky, J. L., Baranowksi, R. L., and Currie, B. L. (1982).Biochem. Biophys. Res. Commun. 109, 1368–1374.
Merrifield, R. B. (1963).J. Am. Chem. Soc. 85, 2149–2154.
Merrifield, R. B., Vizioli, L. D., and Boman, H. G. (1982).Biochemistry 21, 5020–5031.
Mirmira, R. G., and Tager, H. S. (1989).J. Biol. Chem. 264, 6349–6354.
Mirmira, R. G., Nakagawa, S. H., and Tager, H. S. (1991).J. Biol. Chem. 166, 1428–1436.
Mitchell, A. R., Kent, S. B. H., Engelhard, M., and Merrifield, R. B. (1978).J. Org. Chem. 43, 2845–2852.
Nagaraj, R., Sudha, T. S., Shivayi, S., and Balaram, P. (1979).FEBS Lett. 106, 271–274.
Nagaraj, R., and Balaram, P. (1981a).Biochemistry 20, 2828–2835.
Nagaraj, R., and Balaram, P. (1981b).Acc. Chem. Res. 14, 356–382.
Nakagawa, S. H., and Tager, H. S. (1986).J. Biol. Chem. 261, 7332–7341.
Nakagawa, S. H., and Tager, H. S. (1987).J. Biol. Chem. 262, 12,054–12,058.
Ramachandran, J., Berger, A., and Katchalski, E. (1971).Biopolymers 10, 1829–1851.
Schwartz, G. P., and Katsoyannis, P. G. (1973a).J. Chem. Soc. Perkin Trans. 1, 2890–2894.
Schwartz, G. P., and Katsoyannis, P. G. (1973b).J. Chem. Soc. Perkins. Trans. 1, 2894–2901.
Schwartz, G. P., Burke, G. T., Chanley, J. D., and Katsoyannis, P. G. (1983).Biochemistry 22, 4561–4567.
Schwartz, G. P., Burke, G. T., and Katsoyannis, P. G. (1987).Proc. Natl. Acad. Sci. USA 84, 6408–6411.
Stewart, J. M., Knight, M., Paiva, A. C. M., and Paiva, T. (1972). InProgress in Peptide Research, Lande, S. (ed.), Gordon and Breach, New York, pp. 59–64.
Sudha, T. S., and Balaram, P. (1983).Int. J. Peptide Protein Res. 21, 381–388.
Tam, J. P., Heath, W. F., and Merrifeld, R. B. (1983).J. Am. Chem. Soc. 105, 6442–6455.
Turk, J., Needleman, P., and Marshall, G. R. (1976).Mol. Pharmacol. 12, 217–224.
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Wang, Sh., Hu, Sq., Burke, G.T. et al. Insulin analogues with modifications in the β-turn of the B-chain. J Protein Chem 10, 313–324 (1991). https://doi.org/10.1007/BF01025630
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DOI: https://doi.org/10.1007/BF01025630