Summary
Golden hamster, mouse and rat hepatic cadmium metallothioneins (MT) were purified by Sephadex G-75 gel filtration, DEAE-Sephadex A-25 chromatography and activated Thiol-Sepharose 4B affinity chromatography. Metallothioneins were separated by DEAE-Sephadex A-25 chromatography into two forms: MT-1 and MT-2. In mouse and golden hamster liver, MT-1 was the major form. The purified proteins were homogeneous as judged by polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate. In non-denaturing polyacrylamide gel electrophoresis, migration of mouse, rat and golden hamster hepatic metallothioneins were found to be different. Antibodies to mouse hepatic MT-1 was raised in rabbits. The antiserum cross reacted with mouse and hamster MT-1 and MT-2 giving a single precipitin band. Mouse, rat and hamster hepatic MTs are immunologically identical but electrophoretically different. The kidney and pancreatic MTs of rat and golden hamster were purified by Sephadex G-75 gel filtration. They were immunologically distinct. Pancreas MT formed a line of partial identity with hepatic MTs. Kidney MTs form two precipitin band one identical with the pancreatic form and another of complete identity with the hepatic MTs. This indicates the presence of tissue specific MTs.
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References
Kagi JHR, Vallee BL: Metallothionein: A cadmium and Zn containing protein from equine renal cortex. J Biol Chem 235:3460–3465, 1960.
Wagner GJ, Trotter MM: Inducible cadmium binding complexes of cabbage and Tobacco. Plant Physiol 69:804–809, 1982.
Lerch K: Copper metallothionein, a copper binding protein from Neurospora Crassa. Nature 234:368–370, 1980.
Shaikh ZA, Lucis OJ: Induction of cadmium binding proteins. Fed Proc Fed Am Soc Exp Biol 21:298, 1970.
Brady FO, Panemangalore M, Day FA, Fiskin AM, Peterson G: In vivo and ex vivo induction of rat liver metallothionein. In Kagi JHR, Nordberg M (eds) Metallothionein. Experentia Supplement 34:261–271, 1979.
Durnam DM, Palmiter RD: Transcriptional regulation of the mouse metallothionein-1 gene by heavy metals. J Biol Chem 256:5712–5716, 1981.
Failla ML, Cousin RJ: Zinc accumulation and metabolism in primary cultures of adult rat liver cells. Biochim Biophys Acta 543:293–304, 1978.
Mayo KE, Palmiter RD: Glucocorticoid regulation of metallothionein-1 mRNA synthesis in cultured mouse cells. J Biol Chem 256:2621–2624, 1981.
Oh SH, Deagan JT, Whanger PD, Weswig PH: Biological functions of metallothionein V. Am J physiol 234:282–285, 1978.
Sobocinski PZ, Canterbury WJ, Mapes CA: Involvement of hepatic metallothionein, in hypozincemia associated with bacterial infection. Am J Physiol 234:399–406, 1978.
Kagi JHR, Nordberg M: Metallothionein. Experentia Supplement 34:62, 1978.
Whanger PD, Ridlington JW: Role of metallothionein in Zinc metabolism. In Foulkes EC (ed) Biological Roles of Metallothionein. Elsevier North Holland Inc, 1982, pp 264–265.
Otvos JD, Armitage IM: Elucidation of metallothionein structure by 113Cd NMR. In Biochemical Structure Determination by NMR. Marcel Dekker Inc, New York, 1982, pp 66–68.
Karlin M, Richards RI: Human metallothionein genesprimary structure of the metallothionein-II gene and a related processed gene. Nature 299:797–802, 1982.
Kagi JHR, Kojima Y, Kissling MM, Lerch R: Metallothionein: an exceptional metal thiolate protein, CIBA Foundation Symposium-72. Elsevier North Holland Inc, 1980, pp 223–227.
Otvos JD, Olafson RW, Armitage IM: Structure of an invertebrate metallothionein from Scylla serrata. J Biol Chem 257:2427–2431, 1982.
Kagi JHR, Nordberg M: Metallothionein. Experentia Supplement 34:60, 1978.
Maiti I, Mbikay M, Marengo C, Thirion JP: Immunological characterization of metallothioneins in mouse LMTK cells and in a varient, Resistant to Cadmium. J Cell Physiol 112:35–41, 1982.
Fricke U: Tritosal: A new scintillation cocktail based on Triton X-100. Anal Biochem 63:555–558, 1975.
Davis BJ: Disc. Electrophoresis-II Method and application to human serum proteins. Ann NY Acad Sci 121:248–254, 1964.
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, 1970.
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254, 1976.
Vander Mallie RJ, Garvey JS: Radioimmunoassay of metallothioneins. J Biol Chem 254:8416–8421, 1981.
Vander Mallie RJ, Garvey JS: Production and study of antibody produced against rat cadmium thionein. Immunochemistry 15: 857–866, 1978.
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Chakrabarty, T., Maiti, I.B. Immunological and biochemical properties of metallothioneins of golden hamster, mouse and rat. Mol Cell Biochem 68, 41–47 (1985). https://doi.org/10.1007/BF00219387
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DOI: https://doi.org/10.1007/BF00219387