Abstract
The class I and IIClostridium histolyticum collagenases (CHC) have been used to identify hyperreactive sites in rat type I, bovine type II, and human type III collagens. The class I CHC attack both collagens at loci concentrated in the N-terminal half of these collagens starting with the site closest to the N-terminus. The class II CHC initiate collagenolysis by attacking both collagens in the interior to produce a mixture of C-terminal 62,000 and a N-terminal 36,000 fragments. Both fragments are next shortened by removal of a 3000 fragment. These results are very similar to those reported earlier for the hydrolysis of rat type I collagen by these CHC, indicating that the three collagens share many hyperreactive sites. Similar reactions carried out with the respective gelatins show that they are cleaved at many sites at approximately the same rate. Thus, the hyperreactivity of the sites identified must be attributed to their environment in the native collagens. N-terminal sequencing of the fragments produced in these reactions has allowed the identification of 16 cleavage sites in the α1(I), α2(I), α1(II), and α1(III) collagen chains. An analysis of the triple helical stabilities of these cleavage site regions as reflected by their imino acid contents fails to yield a correlation between reactivity and triple helical stability. The existence of these hyperreactive CHC cleavage sites suggests that type I, II, and III collagens contain regions that have specific nontriple helical conformations. The sequence of these sites presented here now makes it possible to investigate these conformations by computational and peptide mimetic techniques.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Angleton, E. L., and Van Wart, H. E. (1988a).Biochemistry 27, 7406–7412.
Angleton, E. L., and Van Wart, H. E. (1988b).Biochemistry 27, 7413–7418.
Bond, M. D., and Van Wart, H. E. (1984a).Biochemistry 23, 3077–3085.
Bond, M. D., and Van Wart, H. E. (1984b).Biochemistry 23, 3085–3091.
Bond, M. D., and Van Wart, H. E. (1984c).Biochemistry 23, 3092–3099.
Chapman, J. A., Holmes, D. F., Meek, K. M., and Rattew, C. J. (1981). InStructural Aspects of Recognition and Assembly in Biological Macromolecules (Balaban, M., Sussman, J. L., Traub, W., and Yonath, A., eds.), Intl. Science Services, Rehovot/Philadelphia, pp. 387–401.
Cozzarelli, N. R., and Wang, J. C. (1990). InDNA Topology and its Biological Effects, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Fasman, G. D. (1977).Handbook of Biochemistry and Molecular Biology: Vol. III, 3rd ed., CRC Press, Cleveland, Ohio, pp. 474–489.
French, M. F., Mookhtiar, K. A., and Van Wart, H. E. (1987).Biochemistry 26, 681–687.
Fujii, K., and Tanzer, M. L. (1976).Biochem. Biophys. Res. Commun. 69, 128–134.
Goldberg, G. J., Wilhelm, S. M., Kronberger, A., Bauer, E. A., Grant, G. A., and Eisen, A. Z. (1986).J. Biol. Chem. 261, 6600–6605.
Gornall, A. G., Bardawill, C. J., and David, M. M. (1949).J. Biol. Chem. 177, 751–766.
Gross, J., Highberger, J. H., Johnson-Wint, B., and Biswas, C. (1980). InCollagenase in Normal and Pathological Tissues (Woolley, D. E., and Evanson, J. M., eds.), John Wiley & Sons, New York, pp. 11–35.
Hasty, K. A., Jeffrey, J. J., Hibbs, M. S., and Welgus, H. G. (1987).J. Biol. Chem. 262, 10,048–10,052.
Hasty, K. A., Pourmotabbed, T. F., Goldberg, G. I., Thompson, J. P., Spinella, D. G., Stevens, R. M., and Mainardi, C. L. (1990).J. Biol. Chem. 265, 11,421–11,424.
Horwitz, A. L., Hance, A. J., and Crystal, R. G. (1977).Proc. Natl. Acad. Sci. USA 74, 897–901.
Laemmli, U. K. (1970).Nature 227, 680–685.
Mallya, S. K., Mookhtiar, K. A., Gao, Y., Brew, K., Dioszegi, M., Birkedal-Hansen, H., and Van Wart, H. E. (1990).Biochemistry 29, 10,628–10,634.
Mayne, R., and Burgeson, R. E. (1987).Structure and Function of Collagen Types, Academic Press, Orlando.
Merril, C. R., Goldman, D., Sedman, S. A., and Ebert, M. H. (1981).Science 211, 1437–1438.
Miller, E. J., and Gay, S. (1982).Methods Enzymol. 82, 3–32.
Mookhtiar, K. A., Grobelny, D., Galardy, R. E., and Van Wart, H. E. (1988).Biochemistry 27, 4299–4304.
Mookhtiar, K. A., Mallya, S. K., and Van Wart, H. E. (1986).Anal. Biochem. 158, 322–333.
Mookhtiar, K. A., Steinbrink, D. R., and Van Wart, H. E. (1985).Biochemistry 24, 6527–6533.
Mookhtiar, K. A., and Van Wart, H. E. (1990).Biochemistry 29, 10,620–10,627.
Mookhtiar, K. A., and Van Wart, H. E. (1991).Matrix (in press).
Moos, M., Jr., Nguyen, N. Y., and Liu, T.-Y. (1988).J. Biol. Chem. 263, 6005–6008.
Piez, K. A. (1984). InExtracellular Matrix Biochemistry (Piez, K. A., and Reddi, A. H., eds.), Elsevier, New York, pp. 1–39.
Seifter, S., and Harper, E. (1971).Enzymes, 3rd ed., Vol. 3, Academic Press, New York, pp. 649–697.
Seyer, J. M., Hasty, K. A., and Kang, A. H. (1989).Eur. J. Biochem. 181, 159–173.
Seyer, J. M., and Kang, A. H. (1981).Biochemistry 20, 2621–2627.
Steinbrink, D. R., Bond, M. D., and Van Wart, H. E. (1985).J. Biol. Chem. 260, 2771–2776.
Stricklin, G. P., Bauer, E. A., Jeffrey, J. J., and Eisen, A. Z. (1977).Biochemistry 16, 1607–1615.
Van Wart, H. E., and Steinbrink, D. R. (1985).Biochemistry 24, 6520–6526.
Welgus, H. G., Jeffrey, J. J., and Eisen, A. Z. (1981).J. Biol. Chem. 256, 9511–9515.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
French, M.F., Bhown, A. & Van Wart, H.E. Identification ofClostridium histolyticum collagenase hyperreactive sites in type I, II, and III collagens: Lack of correlation with local triple helical stability. J Protein Chem 11, 83–97 (1992). https://doi.org/10.1007/BF01025095
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01025095