Abstract
Background
Cathepsin B is a mammalian cysteine protease. The enzyme has been suggested to participate in the patophysiological processes of keratoconus as well as in the corneal response to infectious agents. This study describes the localization of cathepsin B in the rat eye.
Methods
Cathepsin B was identified in rat ocular tissues by Western blotting and immunohistochemistry. Cathepsin B mRNA levels were analyzed in the tissues by quantitative real-time cDNA amplification (QRT-PCR).
Results
Cathepsin B is present in the epithelium, in stromal cells and in the endothelium of the cornea. It is also present in the epithelium lining the ciliary processes, in occasional stromal cells in the iris, in the anterior subcapsular lens epithelium and in various cell types in the retina. At all locations cathepsin B is present in cytoplasmic granules, presumably lysosomes. QRT-PCR analysis detected cathepsin B mRNA in all these tissues in amounts correlating to the immunodetection results, suggesting that the enzyme detected is locally produced.
Conclusions
Cathepsin B is present in several tissues and cell types throughout the rat eye. It is localized to cytoplasmic granules, presumably lysosomes. Our results suggest that it is probably also produced in the same cell types.
Similar content being viewed by others
References
Abrahamson M, Salvesen G, Barrett AJ, Grubb AO (1986) Isolation of six cysteine proteinase inhibitors from human urine. Their physicochemical and enzyme kinetic properties and concentrations in biological fluids. J Biol Chem 261:11282–11289
Assfalg-Machleidt I, Jochum M, Klaubert W, Inthorn D, Machleidt W (1988) Enzymatically active cathepsin B dissociating from its inhibitor complexes is elevated in blood plasma of patients with septic shock and some malignant tumors. Biol Chem Hoppe-Seyler 369 [Suppl]: 263–269
Assfalg-Machleidt I, Jochum M, Nast-Kolb D, Siebeck M, Billing A, Joka T, Rothe G, Valet G, Zauner R, Scheuber HP (1990) Cathepsin B-indicator for the release of lysosomal cysteine proteinases in severe trauma and inflammation. Biol Chem Hoppe Seyler 371 [Suppl]: 211–222
Auricchio F, Mollica L, Ligouri A (1972) Inactivation of thyrosine aminotransferase in neutral homogenates and rat liver slices. Biochem J 129:1131–1138
Barrett AJ (1977) Cathepsin B and other thiol proteases. In: Barrett AJ (ed) Proteinases in mammalian cells and tissues. North-Holland, Amsterdam, 181–208
Barrett AJ, Kirschke H (1981) Cathepsin B, cathepsin H, and cathepsin L. Methods Enzymol 80 (Pt C) : 535–561
Bernstein HG, Reichenbach A, Kirschke H, Wiederanders B (1989) Cell type-specific distribution of cathepsin B and D immunoreactivity within the rabbit retina. Neurosci Lett 27:135–138
Bhat SP (2001) The ocular lens epithelium. Biosci Rep 21:537–563
Bjarnadóttir M, Wulff BS, Sameni M (1998) Intracellular accumulation of the amyloidogenic L68Q variant of human cystatin C in NIH/3T3 cells. Mol Pathol 51:317–326
Burleigh MC, Barrett AJ, Lazarus GS (1974) Cathepsin B1. A lysosomal enzyme that degenerates native collagen. Biochem J 137:387–398
Buttle DJ, Burnett D, Abrahamson M (1990) Levels of neutrophil elastase and cathepsin B activities, and cystatins in human sputum: relationship to inflammation. Scand J Clin Lab Invest 50:509–516
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate- phenol-chloroform extraction. Anal Biochem 162:156–159
Corticchiato O, Cajot J-F, Abrahamson M, Chan SJ, Keppler D, Sordat B (1992) Cystatin C and cathepsin B in human colon carcinoma: expression in cell lines and matrix degradation. Int J Cancer 52:645–652
Coulibaly S, Schwihla H, Abrahamson M, Albini A, Cerni C, Clark JL, Ng KM, Katunuma N, Schlappack O, Glössl J, Mach L (1999) Modulation of the invasive properties of murine squamous carcinoma cells by heterologous expression of cathepsin B and cystatin C. Int J Cancer 83:526–531
De Nanteuil G, Portevin B, Benoist A (2001) Disease-modifying anti-osteoarthritic drugs: current therapies and new prospects around protease inhibition. Farmaco 56:107–112
Dong Z, Katar M, Linebaugh BE, Sloane BF, Berk RS (2001) Expression of cathepsins B, D and L in mouse corneas infected with Pseudomonas aeruginosa. Eur J Biochem 268:6408–1646
Duffy MJ (1996) Proteases as prognostic markers in cancer. Clin Cancer Res 2:613–618
Eisenhauer DA, Berger JJ, Peltier CZ, Taylor A (1988) Protease activities in cultured beef lens epithelial cells peak and then decline upon progressive passage. Exp Eye Res 46:579–590
Felbor U, Dreier L, Bryant RA, Ploegh HL, Olsen BR, Mothes W (2000) Secreted cathepsin L generates endostatin from collagen XVIII. EMBO J 19:1187–1194
Ferrara M, Wojcik F, Rhaissi H, Mordier S, Roux M-P, Bécher D (1990) Gene structure of mouse cathepsin B. FEBS Lett 273:195–199
Ferreras M, Felbor U, Lenhard T, Olsen BR, Delaisse J (2000) Generation and degradation of human endostatin proteins by various proteinases. FEBS Lett 486:247–251
Fröhlich E, Klessen C (2001) Enzymatic heterogeneity of bovine retinal pigment epithelial cells in vivo and in vitro. Graefes Arch Clin Exp Ophthalmol 239:25–34
Fukai N, Eklund L, Marneros AG, Oh SP, Keene DR, Tamarkin L, Niemela M, Ilves M, Li E, Pihlajaniemi T, Olsen BR (2002) Lack of collagen XVIII/endostatin results in eye abnormalities. EMBO J 21:1535–1544
Gorthy WC, Azari P (1987) Biochemical and histochemical evidence for lysosomal proteases in rodent lenses. Exp Eye Res 44:747–753
Guinec N, Dalet-Fumeron V, Pagano M (1993) “In vitro” study of basement membrane degradation by the cysteine proteinases, cathepsins B, B-like and L. Digestion of collagen IV, laminin, fibronectin, and release of gelatinase activities from basement membrane fibronectin. Biol Chem Hoppe Seyler 374:1135–1146
Hahn U, Swanson AA, Hockwin O (1976) Age-related changes in the proteolytic enzymes of mammalian lens. Graefes Arch Klin Exp Ophthalmol 199:197–206
Hayasaka S, Shiono T, Hara S, Fukuyo T (1983) Partial purification of cathepsin B in the bovine ciliary body and iris. Invest Ophthalmol Vis Sci 24:682–686
Illy C, Quraishi O, Wang J, Purisima E, Vernet T, Mort JS (1997) Role of the occluding loop in cathepsin B activity. J Biol Chem 270:1197–1202
Kato T, Chang JH, Azar DT (2003) Expression of type XVIII collagen during healing of corneal incisions and keratectomy wounds. Invest Ophthalmol Vis Sci 44:78–85
Kawabata T, Nishimura Y, Higaki M, Kato K (1993) Purification and processing of rat liver procathepsin B. J Biochem (Tokyo) 113:389–394
Keppler D, Waridel P, Abrahamson M, Bachmann D, Berdoz J, Sordat B (1994) Latency of cathepsin B secreted by human colon carcinoma cells is not linked to secretion of cystatin C and is relieved by neutrophil elastase. Biochim Biophys Acta 1226:117–125
Kos J, Werle B, Lah T, Brunner N (2000) Cysteine proteinases and their inhibitors in extracellular fluids: markers for diagnosis and prognosis in cancer. Int J Biol Markers 15:84–89
Lerch MM, Halangk W, Kruger B (2000) The role of cysteine proteases in intracellular pancreatic serine protease activation. Adv Exp Med Biol 477:403–411
Mach L, Schwihla H, Stuwe K, Rowan AD, Mort JS, Glossl J (1993) Activation of procathepsin B in human hepatoma cells: the conversion into the mature enzyme relies on the action of cathepsin B itself. Biochem J 15; 293:437–442
Mach L, Mort JS, Glossl J (1994) Maturation of human procathepsin B. Proenzyme activation and proteolytic processing of the precursor to the mature proteinase, in vitro, are primarily unimolecular processes. J Biol Chem 269:13030–13035
Mort JS (1998) Cathepsin B. In: Barrett AJ, Rawlings ND, Woessner JF (eds) Handbook of proteolytic enzymes. Academic Press, London, pp 609–617
Nishimura Y, Kawabata T, Kato K (1988) Identification of latent procathepsins B and L in microsomal lumen: characterization of enzymatic activation and proteolytic processing in vitro. Arch Biochem Biophys 261:64–71
Noya O, Alarcon de Noya B, Losada S, Colmenares C, Guzman C, Lorenzo MA, Bermudez H (2002) Laboratory diagnosis of Schistosomiasis in areas of low transmission: a review of a line of research. Mem Inst Oswaldo Cruz 97 [Suppl] 1:167–169
Poole AR, Tiltman KJ, Recklies AD, Stoker TAM (1978) Differences in secretion of the proteinase cathepsin B at the edges of human breast carcinomas and fibroadenomas. Nature 273:545–547
Price G, Tsui J, Unakar NJ (1981) Arylsulfatase—cytochemical localization in lenses of normal and galactose-fed rats. Curr Eye Res 82:567–577
Roughley PJ, Barrett AJ (1977) The degeneration of cartilidge proteoglycans by tissue proteinases. Proteoglycan structure and its susceptibility to proteolysis. Biochem J 167:629–637
San Segundo B, Chan SJ, Steiner DF (1985) Identification of cDNA clones encoding a precursor of rat liver cathepsin B. Proc Natl Acad Sci USA 82:2320–2324
Sexton PS, Cox JL (1997) Inhibition of motility and invasion of B16 melanoma by the over-expression of cystatin C. Melanoma Res 7:97–101
Sherwin T, Brookes NH, Loh IP, Poole CA, Clover GM (2002) Cellular incursion into Bowman’s membrane in the peripheral cone of the keratoconic cornea. Exp Eye Res 74:473–482
Sloane BF, Honn KV (1984) Cysteine proteinases and metastasis. Cancer Metastasis Rev 3:249–263
Swanson AA, Jeter J, Raley KW (1967) Histochemical inentification of lysosomal enzymes in lens epithelial cells. Exp Eye Res 6:351–355
Tsung PK, Lombardini JB (1985) Cathepsin B and D, and Ca2+-dependent neutral protease activities in the retina of taurine-depleted rats. Exp Eye Res 41:285–290
Unakar NJ, Harries W, Tsui J (1985) Acid phosphatase II. Cytochemical localization in lenses of normal and galactose-fed rats. Exp Eye Res 40:117–126
Wassélius J, Håkansson K, Johansson K, Abrahamson M, Ehinger B (2001) Identification and localization of retinal cystatin C. Invest Ophthalmol Vis Sci 42:1901–1906
Zatterstrom UK, Felbor U, Fukai N, Olsen BR (2000) Collagen XVIII/endostatin structure and functional role in angiogenesis. Cell Struct Funct 25:97–101
Zeeuwen PL, van Vlijmen-Willems IM, Hendriks W, Merkx GF, Schalkwijk J (2002) A null mutation in the cystatin M/E gene of ichq mice causes juvenile lethality and defects in epidermal cornification. Hum Mol Genet 11:2867–2875
Acknowledgements
Supported by The Foundation Fighting Blindness, the Knut and Alice Wallenberg Foundation, the Crafoord Foundation, the Swedish Medical Research Council (projects K99-04X, 09915 and 14X-2321) and the Faculty of Medicine at the University of Lund.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wassélius, J., Wallin, H., Abrahamson, M. et al. Cathepsin B in the rat eye. Graefe's Arch Clin Exp Ophthalmol 241, 934–942 (2003). https://doi.org/10.1007/s00417-003-0782-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00417-003-0782-x