Abstract
Transglutaminase (TGase) activities were measured in rat tissues 1-7 days after intraperitoneal injection of saline or lipopolysaccharide (LPS) and in the cells and media from pre-confluent human fibroblasts cultured for two days in the presence or absence of LPS. ε(γ-glutamyl)lysine and [3H]putrescine-labelled γ-glutamyl derivatives in extracellular and cellular fibroblast proteins were also measured. Three effects of LPS were observed. Firstly, total TGase activity is greater in the tissues from the LPS-injected animals, with the maximum increase occurring at 1 day in dermis, epidermis and liver, at 5 days in the aorta and, after a decrease at 2-5 days, at 7 days in the panniculus muscle. Secondly, the fraction of the total activity which is buffer-extractable is greater on days 1 and/or 2 in all the tissues from the LPS-injected rats. Thirdly, in cultures of human fibroblasts, LPS increases that fraction of bound [3H]putrescine and of TGase and its γ-glutamylamine products which occurs in the extracellular medium. In addition, a higher concentration of TGase-derived crosslinks was found in extracellular as opposed to intracellular proteins. In conjunction with previous findings in skin wound healing and in atherosclerosis these results support the concept of an extracellular function for tissue TGase and indicate that there is a widespread association of increases in TGase and its extracellular products with inflammation and the healing or fibrotic processes which follow it.
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References
Lorand L, Conrad SM: Transglutaminases. Mol Cell Biochem 58: 9–35, 1984
Greenberg CS, Birckbichler PJ, Rice RH: Transglutaminases: multi-functional crosslinking enzymes that stabilize tissues. FASEB J 5: 3071–3077, 1991
Aeschlimann D, Paulsson M: Transglutaminases: protein cross-linking enzymes in tissues and body fluids. Thromb Haemost 71: 402–415, 1994
Bowness JM, Tarr AH, Wong T: Increased transglutaminase activity during skin wound healing in rats. Biochim Biophys Acta 967: 234–240, 1988
Wiebe RI, Tarr AH, Bowness JM: Increased transglutaminase in the aortas of cholesterol-fed rabbits: occurrence of buffer-soluble and in-soluble forms and an inhibitor. Biochem Cell Biol 69: 821–827, 1991
Mast BA: The skin. In: IK Cohen, RF Diegelmann and WJ Lindblad (eds). Wound Healing, Biochemical and Clinical Aspects. Saunders, Philadelphia, 1992, pp 344–355
Alexander RW: Inflammation and coronary artery disease. N Engl J Med 331: 468–469, 1994
Leeson MC, Fujihara Y, Morrison DC: Evidence for LPS as the pre-dominant proinflammatory mediator in supernatants of antibiotic-treated bacteria. Infect Immun 62: 4975–4980, 1994
Lowry OH, Rosebrough NH, Farr AL, Randall RJ: Protein measurement with the Folin reagent. J Biol Chem 193: 265–275, 1951
Bowness JM, Venditti M, Tarr AH, Taylor JR: Increase in e(g-glutamyl) lysine crosslinks in atherosclerotic aortas. Atherosclerosis 111: 247–253, 1994
Beninati S, Martinet N, Folk JE: High performance liquid chromatographic method for the determination of e(g-glutamyl)lysine and bis y-glutamyl derivatives of putrescine and spermidine. J Chromatogr 443: 329–335, 1988
Griffin M, Wilson J: Detection of e(g-glutamyl)lysine. Mol Cell Biochem 58: 37–49, 1984
Hirata K, Kaneko A, Ogawa K, Hayasaka H, Onoe T: Effect of endotoxin on rat liver. Analysis of acid phosphatase isozymes in the liver of normal and endotoxin-treated rats. Lab Invest 43: 165–171, 1980
Hewett JA, Roth RA: The coagulation system, but not circulating fibrinogen, contributes to liver injury in rats exposed to lipopolysaccharide from Gram-negative bacteria. J Pharmacol Exp Ther 272: 53–62, 1995
Griffin M, Smith LL, Wynne J: Changes in transglutaminase activity in an experimental model of pulmonary fibrosis induced by paraquat. Br J Exp Pathol 60: 653–661, 1979
Weinberg JB, Pippen AMM, Greenberg CS: Extravascular fibrin formation and dissolution in synovial tissue of patients with osteoarthritis and rheumathoid arthritis. Arthritis Rheum 34: 996–1005, 1991
Suto N, Ikura K, Sasaki R: Expression induced by IL-6 of tissue transglutaminase in human hepatoblastoma Hep G2 cells. J Biol Chem 268: 7469–7473, 1993
Folk JE, Finlayson JS: The e(g-glutamyl)lysine crosslink and the catalytic role of the transglutaminases. Adv Prot Chem 31: 1–133, 1977
Bowness JM, Folk JE, Timpl R: Identification of a substrate site for liver transglutaminase on the aminopropeptide of type III collagen. J Biol Chem 262: 1022–1024, 1987
Bowness JM, Sewell S, Tarr AH: Increased e(g-glutamyl)lysine crosslinking associated with increased protein synthesis in the inner layers of healing skin wounds. Biochim Biophys Acta 1116: 324–330, 1992
Dolynchuk K, Bendor-Samuel R, Bowness JM: Effect of putrescine on tissue transglutaminase activity in wounds: decreased breaking strength and increased fucoprotein solubility. Plast Reconstr Surg 93: 567–573, 1994
Kleman J-P, Aeschlimann D, Paulsson M, van der Rest M: Transglutaminase-catalysed cross-linking of collagen V/XI in A204 rhabdomyosarcoma cells. Biochemistry 34: 13768–13775, 1995
Bowness JM: [ 3H]Fucose incorporation by healing skin wounds and the effect of transglutaminase inhibitors. Canad J Biochem 60: 777–781, 1982
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Bowness, J.M., Tarr, A.H. Increase in transglutaminase and its extracellular products in response to an inflammatory stimulus by lipopolysaccharide. Mol Cell Biochem 169, 157–163 (1997). https://doi.org/10.1023/A:1006846400478
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DOI: https://doi.org/10.1023/A:1006846400478