Abstract
Prolidase [EC 3.4.13.9] is a ubiquitously distributed imidodipeptidase that catalyzes the hydrolysis of C-terminal proline-containing dipeptides. The enzyme plays an important role in the recycling of proline for collagen synthesis and cell growth. Although, the increase in the enzyme activity is correlated with increased rate of collagen turnover, the mechanism by which prolidase is regulated remain largely unknown. In the present study we found that phosphorylation of fibroblast's prolidase may be an underlying mechanism for up regulation of the enzyme activity. Supporting evidence comes from the following observations: (1) immunoprecipitated prolidase was detected as a phosphotyrosine protein as shown by western immunoblot analysis, (2) tyrosine kinase inhibitor – erbstatin induced (in a dose dependent manner) a decrease in prolidase activity in cultured human skin fibroblasts, (3) anti-phosphotyrosine antibody reduced and phosphotyrosine phosphatase 1B antibody (anti-PTP 1B) increased (in a dose dependent manner) the prolidase activity in extract of fibroblast's homogenate, (4) decrease in prolidase activity from collagenase treated or serum starved fibroblasts can be partially prevented by incubating fibroblast's homogenate extract with anti-PTP 1B antibody. These results provide evidence that prolidase is phosphotyrosine enzyme and suggest that the activity of prolidase may be up regulated by the enzyme phosphorylation.
Similar content being viewed by others
References
Myara I, Charpentier C, Lemonnier A: Prolidase and prolidase deficiency. Life Sci 34: 1985–1998, 1984
Mock WL, Green PC, Boyer KD: Specificity and pH dependence for acylproline cleavage by prolidase. J Biol Chem 265: 19600–1965, 1990
Chamson A, Voigtlander V, Myara I, Frey J: Collagen biosynthetic anomalies in prolidase deficiency: Effect of glycyl-L-proline on the degradation of newly synthesized collagen. Clin Physiol Biochem 7: 128–136, 1989
Yaron A, Naider F: Proline-dependent structural and biological properties of peptides and proteins. Crit Rev Biochem Mol Biol 28: 31–81, 1993
Emmerson KS, Phang JM: Hydrolysis of proline dipeptides completely fulfills the proline requirement in a proline-auxotropic Chinese hamster ovary cell line. J Nutr 123: 909–914, 1993
Jackson SH, Dennis AW, Greenberg M: Iminodipeptiduria: A genetic defect in recycling collagen; a method for determining prolidase in erythrocytes. CMA J 113: 759–763, 1975
Albelda SM, Buck CA: Integrins and other cell adhesion molecules. FASEB J 4: 2868–2880, 1990
Bissel M: How does extracellular matrix direct gene expression? J Theor Biol 99: 31–68, 1981
Bolon I, Gouyer V, Devouassoux M, Vandenbunder B, Wernert N, Moro D, Brambilla C, Brambilla E: Expression of c-ets-1, collagenase 1, and urokinase-type plasminogen activator genes in lung carcinomas. Am J Pathol 147: 1298–1310, 1995
Bornstein P, Sage H: Regulation of collagen gene expression. Prog Nucleic Acid Res Mol Biol 37: 67–106, 1989
Carey DJ: Control of growth and differentiation of vascular cells by extracellular matrix. Ann Rev Physiol 53: 161–177, 1991
Pałka JA, Phang JM: Prolidase activity in fibroblasts is regulated by interaction of extracellular matrix with cell surface integrin receptors. J Cell Biochem 67: 166–175, 1997
Burridge K, Fath K, Kelly T, Nuckolls G, Turner C: Focal adhesions: transmembrane junctions between the extracellular matrix and cytoskeleton. Annu Rev Cell Biol 4: 487–525, 1988
Turner CE, Burridge K: Transmembrane molecular assemblies in cellextracellular matrix interactions. Curr Opin Cell Biol 3: 849–853, 1991
Sastry SK, Horowitz AF: Integrin cytoplasmic domains: Mediators of cytoskeletal linkages and extra-and intracellular initiated transmembrane signaling. Curr Opin Cell Biol 5: 819–831, 1993
Boright AP, Scriver CR, Lancaster GA, Choy F: Prolidase deficiency. Biochemical classification of alleles. Am J Hum Genet 44: 731–740, 1989
Myara I, Charpentier C, Lemonnier A: Optimal conditions for prolidase assay by proline colorimetric determination: Application to imidodipeptiduria. Clin Chim Acta 125: 193–205, 1982
Chinard FP: Photometric estimation of proline and ornithine. J Biol Chem 199: 91–95, 1952
Lowry OH, Rosenbregh NI, Far AL, Randall IR: Protein measurement with the folin reagent. J Biol Chem 193: 265–275, 1951
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227: 680–685, 1970
Miltyk W, Karna E, Wołczyński S, Pałka JA: Insulin-like growth factor I-dependent regulation of prolidase activity in cultured human skin fibroblasts. Mol Cell Biochem 189: 177–184, 1998
Ruoslahti E: Control of cell motility and tumor invasion by extracellular matrix interaction. Br J Cancer 66: 239–242, 1992
Endo F, Tanoue A, Nakai H, Indo Y, Titani K, Matsuda I: Primary structure and gene localization of human prolidase. J Biol Chem 264: 4476–4481, 1989
House C, Baldwin GS, Kemp BE: Synthetic peptide substrates for membrane tyrosine protein kinase stimulated by epidermal growth factor. Eur J Biochem 140: 363–367, 1984
Royce PM, Steinmann B: Prolidase deficiency. In: P.M. Royce, B. Steinmann (eds). Connective Tissue and Its Heritable Disorders: Molecular, Genetic and Medical Aspects. Wiley-Liss, New York, 1993, pp 533–548
Freij BJ, Levy HL, Dudin GL, Mutasim D, Deeb M, Der Kaloustian VM: Clinical and biochemical characteristics of prolidase deficiency in siblings. Am J Med Genet 19: 561–571, 1984
Goodman SI, Solomons CC, Muscheinhein F: A syndrome resembling lathyrism associated with iminodipeptiduria. Am J Med 45: 152–159, 1968
Jackson SH, Dennis AW, Greenberg M: Iminodipeptiduria: A genetic defect in recycling of collagen; a method for determining prolidase in erythrocytes. CMA J 113: 759–763, 1975
Fremau RT, Caron MG, Blakely RD: Molecular cloning and expression of a highly affinity L-proline transporter expressed in putative glutamatergic pathways of rat brain. Neuron 8: 915–928, 1992
Miltyk W, Karna E, Pałka J: Inhibition of prolidase activity by nonsteroid antiinflammatory drugs in cultured human skin fibroblasts. Pol J Pharmacol 48: 609–613, 1996
Muszyńska A, Pałka J, Gorodkiewicz E: The mechanism of Daunorubicin-induced inhibition of prolidase activity in human skin fibroblasts and its implication to impaired collagen biosynthesis. Exp Toxic Pathol 52: 149–155, 2000
Pałka J, Miltyk W, Karna E, Wołczyński S: Modulation of prolidase activity during in vitro aging of human skin fibroblasts. The role of extracellular matrix collagen. Tokai J Exp Clin Med 21: 207–213, 1996
Miltyk W, Pałka JA: Potential role of pyrroline 5-carboxylate in regulation of collagen biosynthesis in cultured human skin fibroblasts. Comp Biochem Physiol (part A) 125: 265–271, 2000.
Pałka JA, Karna E, Miltyk W: Fibroblast chemotaxis and prolidase activity modulation by insulin-like growth factor II and mannose 6-phosphate. Mol Cell Biochem 168: 177–183, 1997
Senboshi Y, Oono T, Arata J: Localization of prolidase gene expression in scar tissue using in situ hybridization. J Dermatol Sci 12: 163–171, 1996
Romanic AM, Graesser D, Baron JL, Visintin I, Janeway CA Jr, Madri JA: T cell adhesion to endothelial cells and extracellular matrix is modulated upon transendothelial cell migration. Lab Invest 76: 11–23, 1997
Hauzenberger D, Klominek J, Holgersson J, Bergstrom SE, Sundqvist KG: Triggering of motile behaviour in T lymphocytes via cross-linking of α4β1 and αLβ2. J Immunol 158: 76–84, 1997
Keely PJ, Parise LV: The α2β1 integrin is a necessary co-receptor for collagen-induced activation of Syk and the subsequent phosphorylation of phospholipase C gamma2 in platelets. J Biol Chem 271: 26668–26676, 1996
Boudreau NJ, Jones PL: Extracellular matrix and integrin signalling: The shape of things to come. Biochem J 339: 481–488, 1999
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Surażyński, A., Pałka, J. & Wołczyński, S. Phosphorylation of prolidase increases the enzyme activity. Mol Cell Biochem 220, 95–101 (2001). https://doi.org/10.1023/A:1010849100540
Issue Date:
DOI: https://doi.org/10.1023/A:1010849100540