β-Glucuronidase of Rat Preputial Gland1

Himeno, Masaru; Ohhara, Hideto; Arakawa, Yasuaki; Kato, Keitaro

doi:10.1093/oxfordjournals.jbchem.a130742

Cited by 55 publications

(20 citation statements)

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“…fl-glucuronidase was purified to homogeneity from rat preputial glands (11). Mannose-bovine serum albumin (Man-BSA) was prepared as described elsewhere (12) and contained 20-40 mol of sugar per ml of protein.…”

Section: Methodsmentioning

confidence: 99%

Receptor-mediated entry of beta-glucuronidase into the parasitophorous vacuoles of macrophages infected with Leishmania mexicana amazonensis.

Shepherd¹,

Stahl²,

Bernd³

et al. 1983

The Journal of Experimental Medicine

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125I-labeled rat preputial gland beta-glucuronidase was shown by light and electron microscopic radioautography to accumulate within the parasitophorous vacuoles of in vitro derived bone marrow macrophages infected with Leishmania mexicana amazonensis. beta-glucuronidase uptake was mediated by the mannose receptor, since the penetration of the ligand was inhibited by mannan. Uptake was detected as soon as 4 h after incubation of infected cells with the ligand, and increased at 24 and 48 h. The label persisted in the vacuoles for at least 24 h after a 24-h pulse with the ligand, a finding compatible with the relatively long half-life of labeled beta-glucuronidase in normal macrophages. Parasitophorous vacuoles were also labeled in macrophages exposed to the ligand only before infection, indicating that secondary lysosomes containing the ligand fused with the parasitophorous vacuoles. Another mannosylated ligand, mannose-BSA, which, in contrast to beta-glucuronidase, is rapidly degraded in macrophage lysosomes, did not detectably accumulate in the vacuoles. The results support and extend information previously obtained with electron opaque tracers that emphasizes the phagolysosomal nature of Leishmania parasitophorous vacuoles. In addition, the results suggest that appropriate mannosylated molecules may be used as carriers for targeting of leishmanicidal drugs to the parasitophorous vacuoles of infected macrophages.

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Section: Methodsmentioning

confidence: 99%

Receptor-mediated entry of beta-glucuronidase into the parasitophorous vacuoles of macrophages infected with Leishmania mexicana amazonensis.

Shepherd¹,

Stahl²,

Bernd³

et al. 1983

The Journal of Experimental Medicine

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“…Glycoconjugates of milk sugar oligosaccharides and bovine serum albumin were prepared by the method of Zopf et aL (19). (-Glucuronidase was purified to homogeneity from rat preputial glands by the method of Himeno et aL (20). The enzyme was assayed by using phenolphthalein glucuronide as described by Stahl and Touster (21).…”

Section: Methodsmentioning

confidence: 99%

L-Fucose-terminated glycoconjugates are recognized by pinocytosis receptors on macrophages.

Shepherd¹,

Lee²,

Schlesinger³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

166

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'MI-Labeled L-fucose-albumin complex and rat preputial fi-glucuronidase are rapidly cleared from plasma after intravenous infusion. L-Fucose-albumin retards the plasma clearance of fi-glucuronidase whereas D-fucose-albumin is inactive. In vitro, '5I-labeled L-fucose-albumin is taken up into rat or rabbit alveolar macrophages by receptor-mediated pinocytosis. Uptake (37°C) is time-dependent, is saturable with increasing ligand concentration (KY e = 4.4 X 10-8M), and requires Ca2". i25ILabeled D-fucose-abumin is poorly taken up. Binding (4°C) is saturable and Ca2+ dependent. Binding and uptake are fully inhibited by yeast mannan. A series of neoglycoproteins, including Lfucose-albumin, were tested as inhibitors of uptake of '25I-labeled 3-glucuronidase into macrophages. The following order of potency was observed:L-Fucose-terminated oligosaccharides coupled to bovine serum albumin also block`5I-labeled (3-glucuronidase uptake into macrophages.

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“…2 and 3). Since the molecular mass of the B-glucuronidase subunit is -75 kDa (4), the open reading frame within the two overlapping clones must represent -50% of the p-glucuronidase mRNA coding region. Indeed, the size of the preputial gland mRNA encoding ,3-glucuronidase was shown to be 2.6 kb by RNA blot analysis, using as a probe a 32P-labeled plasmid containing the recombined p,BG2 and pPG5 inserts (Fig.…”

Section: Imentioning

confidence: 99%

“…The richest known source of f3-glucuronidase is the preputial gland of female rats, where this enzyme comprises =7% of the total protein (3). Rat preputial gland ,-glucuronidase is a tetrameric glycoprotein composed of identical subunits with a molecular mass of -75 kDa (4).…”

mentioning

confidence: 99%

Nucleotide sequence of rat preputial gland beta-glucuronidase cDNA and in vitro insertion of its encoded polypeptide into microsomal membranes.

Nishimura¹,

Rosenfeld²,

Kreibich³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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We have selected the rat preputial gland I3-glucuronidase as a model protein to study the sorting of newly synthesized lysosomal hydrolases to the lysosome. The complete coding sequence of j3-glucuronidase messenger RNA was determined from the sequences of a group of overlapping cDNA clones isolated from preputial gland cDNA libraries. The fi-glucuronidase mRNA primary translation product contains 648 amino acids, including an amino-terminal signal sequence of 22 residues. The polypeptide has four potential sites for the addition of asparagine-linked core oligosaccharides. A 376-residue segment of fi-glucuronidase shows extensive homology (23% sequence identity) to a portion of Escherichia coli .l-galactosidase. This homology most likely reflects an evolutionary relationship between the bacterial and eukaryotic enzymes and the conservation of structural features necessary for the glycosidase activity of both proteins. Translation of mRNA synthesized in vitro by transcription of a cDNA containing the entire ,B-glucuronidase coding region yielded a polypeptide that was immunoprecipitated with anti-,B-glucuronidase antiserum and had the same electrophoretic mobility as the primary translation product of natural ,B-glucuronidase mRNA. In the presence of microsomal membranes, the in vitro-synthesized (3-glucuronidase underwent cotranslational incorporation into the microsomes, as indicated by removal of the signal sequence and the addition of several oligosaccharide chains. The Pglucuronidase cDNA will provide a useful tool to study the mechanism of mannose phosphorylation and other aspects of

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β-Glucuronidase of Rat Preputial Gland1

Cited by 55 publications

References 0 publications

Receptor-mediated entry of beta-glucuronidase into the parasitophorous vacuoles of macrophages infected with Leishmania mexicana amazonensis.

Receptor-mediated entry of beta-glucuronidase into the parasitophorous vacuoles of macrophages infected with Leishmania mexicana amazonensis.

L-Fucose-terminated glycoconjugates are recognized by pinocytosis receptors on macrophages.

Nucleotide sequence of rat preputial gland beta-glucuronidase cDNA and in vitro insertion of its encoded polypeptide into microsomal membranes.

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