Collagenase from corneal cell cultures and its modulation by phagocytosis.

The uptake of latex by fibroblasts in confluent primary culture results in the secretion of collagenase at a linear rate for a prolonged period. Phagocytosis might therefore constitute an important level of collagenase regulation in corneal ulceration. The collagenase in cell cultures is present in a latent form (40,000 MW) like that obtained from organ cultures of ulcerating corneas and can be activated proteolytically. Production of the latent collagenase in cell culture depends upon the presence of serum and diminishes greatly when serum is removed from the medium. Collagenase activity can be demonstrated after the latent collagenase has been separated from serum antiproteases in the media. Alternatively, careful titration of the crude media with trypsin to saturate serum antiproteases, to release collagenase from the complex with alpha 2-macroglobulin, and to activate latent collagenase also results in measurable collagenase activity. The collagenase that is secreted cleaves fibrillar type I collagen and cleaves soluble type I collagen into the typical 3/4 and 1/4 length fragments, as demonstrated by SDS-gel electrophoresis and electron microscopy.

[1]  K. Kenyon,et al.  Prevention of stromal ulceration in the alkali-burned rabbit cornea by glued-on contact lens. Evidence for the role of polymorphonuclear leukocytes in collagen degradation. , 1979, Investigative ophthalmology & visual science.

[2]  J. Gross,et al.  Regulation of corneal collagenase production: epithelial-stromal cell interactions. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[3]  M. Berman,et al.  Latent collagenase in the ulcerating rabbit cornea. , 1977, Experimental eye research.

[4]  T. Sun,et al.  Cultured epithelial cells of cornea, conjunctiva and skin: absence of marked intrinsic divergence of their differentiated states , 1977, Nature.

[5]  M. Berman,et al.  5' Adenosine memophosphate prevents collagen degradation in culture but does not prevent corneal ulceration. , 1977, Experimental eye research.

[6]  H. Birkedal‐Hansen,et al.  Synthesis and release of procollagenase by cultured fibroblasts. , 1976, The Journal of biological chemistry.

[7]  D. Woolley,et al.  Small molecular weight β1 serum protein which specifically inhibits human collagenases , 1976, Nature.

[8]  H. D. Cavanagh,et al.  Regulation of collagenase activity in the ulcerating cornea by cyclic-AMP. , 1976, Experimental eye research.

[9]  H. Birkedal‐Hansen,et al.  Activation of latent bovine gingival collagenase. , 1975, Archives of oral biology.

[10]  A. Eisen,et al.  Collagenase production by human skin fibroblasts. , 1975, Biochemical and biophysical research communications.

[11]  Z. Werb,et al.  STIMULATION BY ENDOCYTOSIS OF THE SECRETION OF COLLAGENASE AND NEUTRAL PROTEINASE FROM RABBIT SYNOVIAL FIBROBLASTS , 1974, The Journal of experimental medicine.

[12]  D. Deporter,et al.  The role of the fibroblast in collagen turnover in the functioning periodontal ligament of the mouse. , 1974, Archives of oral biology.

[13]  Z. Werb,et al.  A specific collagenase from rabbit fibroblasts in monolayer culture. , 1974, The Biochemical journal.

[14]  M. Berman,et al.  Corneal collagenases: evidence for zinc metalloenzymes. , 1973, Annals of ophthalmology.

[15]  S. I. Brown,et al.  Fibroblast collagenase partial purification and characterization. , 1973, Investigative ophthalmology.

[16]  P. Davison,et al.  Tissue collagenase: a simplified, semiquantitative enzyme assay. , 1973, Analytical biochemistry.

[17]  A. Barrett,et al.  The interaction of α2-macroglobulin with proteinases. Characteristics and specificity of the reaction, and a hypothesis concerning its molecular mechanism , 1973 .

[18]  P. Davison,et al.  Characterization of human corneal collagenase. , 1973, Experimental eye research.

[19]  P. Robertson,et al.  Collagenase: Localization in Polymorphonuclear Leukocyte Granules in the Rabbit , 1972, Science.

[20]  M. Glimcher,et al.  Maintenance of Mouse Bone Collagenase Activity in the Presence of Serum Protein by Addition of Trypsin 1 , 1972, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[21]  S. I. Brown,et al.  Pathogenesis of ulcers of the alkali-burned cornea. , 1970, Archives of ophthalmology.

[22]  C. Dohlman,et al.  Collagenase in the cornea. , 1969, Experimental eye research.

[23]  Weimar,et al.  POLYMORPHONUCLEAR INVASION OF WOUNDED CORNEAS , 1957, The Journal of experimental medicine.

[24]  Berman Mb,et al.  The role of alpha-macroglobulins in corneal ulceration. , 1976, Progress in clinical and biological research.

[25]  J. Gross.  Aspects of the Animal Collagenases , 1976 .

[26]  P. C. Harpel.  Human alpha2-macroglobulin. , 1976, Methods in enzymology.

[27]  M. Berman.  Collagenase Inhibitors: Rationale for Their Use in Treating Corneal Ulceration , 1975, International ophthalmology clinics.

[28]  C. Dohlman,et al.  Collagenase inhibitors. , 1975, Archives d'ophtalmologie et revue generale d'ophtalmologie.

[29]  H. D. Cavanagh,et al.  Herpetic ocular disease: therapy of persistent epithelial defects. , 1975, International ophthalmology clinics.

[30]  H. Coon.  Clonal stability and phenotypic expression of chick cartilage cells in vitro. , 1966, Proceedings of the National Academy of Sciences of the United States of America.