Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme

von Clausbruch UC, Tschesche H (1988)
Biol Chem Hoppe Seyler 369(8): 683-691.

Journal Article | Published | English

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Cathepsin D of human leukocytes was isolated and characterized. Purified leukocytes were lysed under nitrogen pressure and the proteinase activity precipitated by centrifugation at 48,000 x g. The precipitate was extracted by various buffers. The yield of cathepsin D was almost pH-independent but could be increased by Triton X-100. Employing gel chromatography the activity was found at a molecular mass close to 42,000 Da. Purification of the enzyme was performed by a two-step procedure using pepstatin-Sepharose chromatography and ion exchange chromatography. Three multiple forms of the enzyme were separated by ion exchange chromatography. The isoelectric points of the three forms of the enzyme were close to pH 5.0. The enzyme showed the typical characteristics of the acid proteinase cathepsin D. Enzyme activity was influenced by heavy metals such as Hg2 and Fe3 as well as by typical inhibitors for carboxyl-proteinases such as diazoacetyl-DL-norleucine methyl ester, 1,2-epoxy-3-(4-nitrophenoxy)propane and 4-bromo-phenacylbromide. An immunological comparison with cathepsin D from human liver by immunodiffusion and immunoelectrophoresis indicates identity of the two enzymes.
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von Clausbruch UC, Tschesche H. Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme. Biol Chem Hoppe Seyler. 1988;369(8):683-691.
von Clausbruch, U. C., & Tschesche, H. (1988). Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme. Biol Chem Hoppe Seyler, 369(8), 683-691.
von Clausbruch, U. C., and Tschesche, H. (1988). Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme. Biol Chem Hoppe Seyler 369, 683-691.
von Clausbruch, U.C., & Tschesche, H., 1988. Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme. Biol Chem Hoppe Seyler, 369(8), p 683-691.
U.C. von Clausbruch and H. Tschesche, “Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme”, Biol Chem Hoppe Seyler, vol. 369, 1988, pp. 683-691.
von Clausbruch, U.C., Tschesche, H.: Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme. Biol Chem Hoppe Seyler. 369, 683-691 (1988).
von Clausbruch, U C, and Tschesche, Harald. “Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme”. Biol Chem Hoppe Seyler 369.8 (1988): 683-691.
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PMID: 3145753
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