Biopolym. Cell. 2000; 16(6):487-494.
Structure and Function of Biopolymers
Properties of high-molecular-mass ATP-dependent proteinase from the erythroid cells of piglets
1Antonyak H. L.
  1. Institute of Animal Biology, NAAS of Ukraine
    38, Stusa Str., Lviv, Ukraine, 79034

Abstract

The present article reports the characterization of high-molecular-mass ATP-dependent proteinase from the erythroid ceils of neonatal piglets. The enzyme was purified by DEAE-Toyopearl 650M, Toyopearl HW-55 gel filtration and hydroxyapatite chromato-graphies. The molecular mass of enzyme has been found to be 700±50 kDa by gel filtration, the optimum activity was observed at around pH 8.5. The enzyme hydrolysed synthetic substrate Suc-Leu-Leu-Val-Tyr-MCA and showed proteolytic activity towards casein and several o ther proteins in the presence of ATP. The degradation of the protein substrates was stimulated by 0.05 % sodium dodecyl sulfate and linoleic acid. The casein-hydrolyzing activity decreased in the presence of phenylmethylsulfonylfluoride and n-chloromer-curibenzoic acid. T he enzyme activity reached high levels in the proliferating erythroblasts of animal bone marrow and decreased as the cells matured. The high levels of the proteinase activity were observed in the erythroid cells of newborn piglets, whereas in the cells of 10-days-old animals the enzyme activity was significantly lower. A role of ATP-dependent high-molecular-mass proteinase in functional activity of animal erythroid cells in the neonatal period discussed.

References

[1] Bond JS, Butler PE. Intracellular proteases. Annu Rev Biochem. 1987;56:333-64.
[2] Sologub LI, Pashkovska IS, Antonyak GL. Cells proteases and their functions. K.: Naukova Dumka, 1992; 194 p.
[3] Jarett L, Kiechle FL, Parker JC. Chemical mediator or mediators of insulin action: response to insulin and mode of action. Fed Proc. 1982;41(11):2736-41.
[4] Solohub LI, Pashkovsk'ka IS, Sukhors'ka IE, Antoniak HL. [Role of proteolysis in posttranslational modifications of biologically active peptides and polypeptides]. Ukr Biokhim Zh. 1991;63(5):3-14.
[5] Solohub LI, Pashkovskaya IS, Antoniak GL. Participation of proteinases in invasion of malignant tumours. Exp Oncol. 1992; 14(6):7-13.
[6] Rivett AJ. High molecular mass intracellular proteases. Biochem J. 1989;263(3):625-33.
[7] Gronostajski RM, Pardee AB, Goldberg AL. The ATP dependence of the degradation of short- and long-lived proteins in growing fibroblasts. J Biol Chem. 1985;260(6):3344-9.
[8] Dahlmann B, Kopp F, Kuehn L, Niedel B, Pfeifer G, Hegerl R, Baumeister W. The multicatalytic proteinase (prosome) is ubiquitous from eukaryotes to archaebacteria. FEBS Lett. 1989;251(1-2):125-31.
[9] Orlowski M. The multicatalytic proteinase complex, a major extralysosomal proteolytic system. Biochemistry. 1990;29(45):10289-97.
[10] Reits EA, Benham AM, Plougastel B, Neefjes J, Trowsdale J. Dynamics of proteasome distribution in living cells. EMBO J. 1997;16(20):6087-94.
[11] Hicke L. Ubiquitin-dependent internalization and down-regulation of plasma membrane proteins. FASEB J. 1997;11(14):1215-26.
[12] Palombella VJ, Rando OJ, Goldberg AL, Maniatis T. The ubiquitin-proteasome pathway is required for processing the NF-kappa B1 precursor protein and the activation of NF-kappa B. Cell. 1994;78(5):773-85.
[13] Peters JM. Proteasomes: protein degradation machines of the cell. Trends Biochem Sci. 1994;19(9):377-82.
[14] Harris JR. Some high molecular-weight oligomeric proteins and enzymes of reticulocytes and erythrocytes. Blood Cell Biochemistry. Ed. J. R. Harris. New York: Plenum press, 1990. Vol. 1: 251-98.
[15] Antonyak GL, Snitynskiy VV, Solohub LI, Vernikovska YaI. Studying the properties of macromolecular ATP-dependent proteases erythroid cells pigs. Theses. Intern. sympoz. "Biological mechanisms of aging». Kharkiv, 1996;13.
[16] Starodub NF, Nazarenko VI. Heterogeneous hemoglobin system. K.: Naukova Dumka, 1987; 199 p.
[17] Harrison FL, Beswick TM, Chesterton CJ. Separation of haemopoietic cells for biochemical investigation. Preparation of erythroid and myeloid cells from human and laboratory-animal bone marrow and the separation of erythroblasts according to their state of maturation. Biochem J. 1981;194(3):789-96.
[18] Sizova NA, Kamenskaya VV, Fedenkov VI, Kozarenko NM. Method o frctioning of red n;lood cells in the debsity gradient of saccarose with out usin apparatus. . Izv Sib Otdel Akad NAuk SSSR. 1980; 3(15): 119-22.
[19] Beutler E, West C, Blume KG. The removal of leukocytes and platelets from whole blood. J Lab Clin Med. 1976;88(2):328-33.
[20] Rapoport S, Dubiel W, M?ller M. Proteolysis of mitochondria in reticulocytes during maturation is ubiquitin-dependent and is accompanied by a high rate of ATP hydrolysis. FEBS Lett. 1985;180(2):249-52.
[21] Ishiura S, Sano M, Kamakura K, Sugita H. Isolation of two forms of the high-molecular-mass serine protease, ingensin, from porcine skeletal muscle. FEBS Lett. 1985;189(1):119-23.
[22] Hough R, Pratt G, Rechsteiner M. Purification of two high molecular weight proteases from rabbit reticulocyte lysate. J Biol Chem. 1987;262(17):8303-13.
[23] Tanaka K, Ii K, Ichihara A, Waxman L, Goldberg AL. A high molecular weight protease in the cytosol of rat liver. I. Purification, enzymological properties, and tissue distribution. J Biol Chem. 1986;261(32):15197-203.
[24] Wilk S, Orlowski M. Evidence that pituitary cation-sensitive neutral endopeptidase is a multicatalytic protease complex. J Neurochem. 1983;40(3):842-9.
[25] Dahlmann B, Kuehn L, Rutschmann M, Reinauer H. Purification and characterization of a multicatalytic high-molecular-mass proteinase from rat skeletal muscle. Biochem J. 1985;228(1):161-70.
[26] Coux O, Tanaka K, Goldberg AL. Structure and functions of the 20S and 26S proteasomes. Annu Rev Biochem. 1996;65:801-47.
[27] McDonald JK. An overview of protease specificity and catalytic mechanisms: aspects related to nomenclature and classification. Histochem J. 1985;17(7):773-85.
[28] Dahlmann B, Rutschmann M, Kuehn L, Reinauer H. Activation of the multicatalytic proteinase from rat skeletal muscle by fatty acids or sodium dodecyl sulphate. Biochem J. 1985;228(1):171-7.
[29] Wessels JMC, Veerkamp JH. Some aspects of the osmotic lysis of erythrocytes III. Comparison of glycerol permeability and lipid composition of red blood cell membranes from eight mammalian species. Biochim Biophys Acta. 1973;291(1):190–6.
[30] Shimbara N, Orino E, Sone S, Ogura T, Takashina M, Shono M, Tamura T, Yasuda H, Tanaka K, Ichihara A. Regulation of gene expression of proteasomes (multi-protease complexes) during growth and differentiation of human hematopoietic cells. J Biol Chem. 1992;267(25):18100-9.
[31] Kumatori A, Tanaka K, Inamura N, Sone S, Ogura T, Matsumoto T, Tachikawa T, Shin S, Ichihara A. Abnormally high expression of proteasomes in human leukemic cells. Proc Natl Acad Sci U S A. 1990;87(18):7071-5.
[32] Antoniak GL. [The effect of thyroxine and insulin on the hemopoiesis in animals during neonatal development]. Tsitologiia. 1999;41(6):512-5.
[33] Antoniak GL, Snitynskiy VV, Danchuk VV, Babich YaO, Iskra RYa, Balkovskiy VV, Krektun BV, Buchko OM. Effect of exogenous hormones on functional activity of endocrine glands in neonatal piglets ontogeny. Visn. Bilotserkiv. derzh. ahrar. un-tu. 1998; 5(1):150-3.
[34] Girard J, Ferre P. Metabolic and hormonal changes around birth. Biochemical Development of the Fetus and Neonate. Ed. C. T. Jones. New York etc.: Elsevier, 1982: 517-51.
[35] Biochemical development of the fetus and neonate. Ed. C. T. Jones. New York etc.: Elsevier, 1982. 685 p.
[36] Antonyak GL. The role of the proteolytic enzymes in the functional activity of neutrophils. Usp Sovrem Biol. 1999; 119(5): 475-85.
[37] Harris JR. Some high molecular-weight oligomeric protems and enzymes of reticulocytes and elythrocytes. Blood Cell Biochemistry. Ed. J. R. Harris. New York, etc.: Plenum Press, 1990. V. 1-P. 251-298
[38] Hershko A, Ciechanover A. The ubiquitin system for protein degradation. Annu Rev Biochem. 1992;61:761-807.
[39] Bush KT, Goldberg AL, Nigam SK. Proteasome inhibition leads to a heat-shock response, induction of endoplasmic reticulum chaperones, and thermotolerance. J Biol Chem. 1997;272(14):9086-92.
[40] Ciechanover A, Schwartz AL. The ubiquitin-mediated proteolytic pathway: mechanisms of recognition of the proteolytic substrate and involvement in the degradation of native cellular proteins. FASEB J. 1994;8(2):182-91.