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			PubMed Journals: J Biol Chem

  Source:		PMID: 2168407


    		J Biol Chem. 1990 Sep 5;265(25):15341-5.
     
			Identification of membrane anchoring site
			of human renal dipeptidase and construction
			and expression of a cDNA for its secretory
			form.

			Adachi H(1), Katayama T, Inuzuka C, Oikawa
			S, Tsujimoto M, Nakazato H.

			Author Information
			(1) Suntory Institute for Biomedical Research,
			Osaka, Japan.

			The chemical properties of human renal dipeptidase
			(hrDP) purified from the membrane fraction
			of kidney have been characterized. When
			treated with phosphatidylinositol-specific
			phospholipase C, hrDP was released from
			renal membrane fractions. After digestion
			with trypsin, carboxyl-terminal peptide was isolated
			employing anhydrotrypsin-agarose column
			chromatography and reversed-phase high performance
			liquid chromatography. The amino acid sequence
			of the peptide was identified at positions
			363-369 in the primary structure deduced
			from the cDNA sequence (Adachi, H., Tawaragi,
			Y., Inuzuka, C., Kubota, I., Tsujimoto,
			M., Nishihara, T., And Nakazato, H. (1990)
			J. Biol. Chem. 265, 3992-3995). Further
			examination of the chemical composion of
			the peptide showed that it contained, respectively,
			2, 1, 5, 1, and 1 mol of ethanolamine, glucosamine,
			mannose, inositol, and phosphate in addition
			to amino acids. These results suggest that
			the mature hrDP molecule lacks the carboxyl-terminal
			hydrophobic peptide extension predicted
			from the cDNA sequence and is anchored at
			Ser369 via glycosylphosphatidylinositol
			to the membrane. To characterize further
			the action of the enzyme, we have established
			expression systems for both secretory and
			membrane anchored forms of hrDP using COS-1
			cells and found that both recombinant forms
			were as active as natural enzyme. Our expression
			system made it possible to prepare large
			amounts of soluble enzyme, and will contribute
			toward elucidation of the physiological roles
			of the enzyme.

			PMID: 2168407 [Indexed for MEDLINE]

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