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PMID: 3667691 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Lumenal location of the microsomal beta-glucuronidase-egasyn complex.

The Journal of cell biology ·Vol. 105 ·No. 4 ·1987-10-00 ·Pages 1571-8

Brown J, Novak EK, Takeuchi K, Moore K, Medda S, Swank RT

Abstract

Mouse liver beta-glucuronidase is stabilized within microsomal vesicles by complexation with the accessory protein egasyn. The location of the beta-glucuronidase-egasyn complex and free egasyn within microsomal vesicles was investigated. Surprisingly, it was found that neither the complex nor free egasyn are intrinsic membrane components. Rather, both are either free within the vesicle lumen or only weakly bound to the inside of the vesicle membrane. This conclusion was derived from release studies using low concentrations of Triton X-100 or controlled sonication. Both the intact complex and free egasyn were released in parallel with lumenal proteins, not with intrinsic membrane components. Also, beta-glucuronidase was protected from digestion by proteinase K by the membrane of microsomal vesicles. The hydrophilic nature of both the complex and free egasyn was confirmed by phase separation experiments with the detergent Triton X-114. Egasyn is one of an unusual group of esterases that, despite being located within the lumen or only weakly bound to the lumenal surface of the endoplasmic reticulum, do not enter the secretory pathway.

MeSH Terms
Animals Carboxylic Ester Hydrolases Cell Compartmentation Female Glucuronidase/metabolism Kidney/enzymology,ultrastructure Macromolecular Substances Male Membrane Glycoproteins/metabolism Mice Microsomes/enzymology Microsomes, Liver/enzymology Polyethylene Glycols Solubility
Chemicals
Macromolecular Substances Membrane Glycoproteins Polyethylene Glycols Carboxylic Ester Hydrolases egasyn Glucuronidase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Brown J
Washington State Patrol Crime Lab, Seattle 98104.
Novak E K
Takeuchi K
Moore K
Medda S
Swank R T
References (43)
43 references, click to expand
  1. Procedure for the selective release of content from microsomal vesicles without membrane disassembly.
    Methods Enzymol. 1974;31:215-25 PMID: 4419291
  2. THE CYTOLOGIC DEMONSTRATION OF BETA-GLUCURONIDASE EMPLOYING NAPHTHOL AS-BI GLUCURONIDE AND HEXAZONIUM PARAROSANILIN; A PRELIMINARY REPORT.
    J Histochem Cytochem. 1964 Apr;12:293-7 PMID: 14187340
  3. Guinea pig liver microsomal UDP-glucuronyltransferase: compartmented or phospholipid-constrained?
    Biochim Biophys Acta. 1975 Oct 22;403(2):335-44 PMID: 810164
  4. Defective lysosomal enzyme secretion in kidneys of Chediak-Higashi (beige) mice.
    J Cell Biol. 1975 Dec;67(3):774-88 PMID: 408
  5. Distribution of constitutive enzymes and phospholipids in microsomal membranes of rat liver.
    FEBS Lett. 1975 Oct 15;58(1):190-3 PMID: 817936
  6. Purification and characterization of rat liver microsomal beta-glucuronidase.
    Biochim Biophys Acta. 1976 Jul 8;438(2):474-86 PMID: 8108
  7. Isolation, characterization, and radioimmunoassay of murine egasyn, a protein stabilizing glucuronidase membrane binding.
    J Biol Chem. 1976 Dec 25;251(24):7753-60 PMID: 826534
  8. Turnover of murine beta-glucuronidase. Comparison among liver, kidney, and spleen and between lysosomes and microsomes.
    J Biol Chem. 1978 Aug 10;253(15):5437-42 PMID: 670206
  9. Genetic regulation of mammalian glucuronidase.
    Recent Prog Horm Res. 1978;34:401-36 PMID: 366684
  10. Biosynthesis of two lysosomal enzymes in macrophages. Evidence for a precursor of beta-galactosidase.
    J Biol Chem. 1979 Oct 25;254(20):9939-42 PMID: 114527
  11. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  12. Phase separation of integral membrane proteins in Triton X-114 solution.
    J Biol Chem. 1981 Feb 25;256(4):1604-7 PMID: 6257680
  13. Genetic determination of kinetic parameters in beta-glucuronidase induction by androgen.
    J Biol Chem. 1981 Mar 25;256(6):3005-11 PMID: 7204385
  14. The synthesis and processing of beta-glucuronidase in normal and egasyn deficient mouse kidney.
    Biochem Biophys Res Commun. 1981 Mar 31;99(2):691-9 PMID: 7236294
  15. Turnover of two lysosomal enzymes in macrophages.
    J Biol Chem. 1981 Oct 10;256(19):10137-44 PMID: 6792196
  16. Comparative studies of asparagine-linked oligosaccharide structures of rat liver microsomal and lysosomal beta-glucuronidases.
    Arch Biochem Biophys. 1981 Jun;209(1):298-303 PMID: 6792990
  17. Posttranslational enzymes in the biosynthesis of collagen: intracellular enzymes.
    Methods Enzymol. 1982;82 Pt A:245-304 PMID: 6210830
  18. Nonspecific esterases of Mus musculus.
    Biochem Genet. 1982 Jun;20(5-6):585-606 PMID: 7115289
  19. Interstrain variation of esterase-22, a new isozyme of the house mouse.
    Comp Biochem Physiol B. 1982;73(3):719-24 PMID: 7151411
  20. Carboxyl-terminal proteolytic processing during biosynthesis of the lysosomal enzymes beta-glucuronidase and cathepsin D.
    Biochemistry. 1983 Oct 25;22(22):5201-5 PMID: 6360205
  21. Purification and characterization of various esterases from rat liver.
    Eur J Biochem. 1983 Dec 1;137(1-2):293-301 PMID: 6653557
  22. Preparation of microsomal beta-glucuronidase and its membrane anchor protein, egasyn.
    Methods Enzymol. 1983;96:557-69 PMID: 6656645
  23. Isolation of a homogeneous glucosidase II from pig kidney microsomes.
    Eur J Biochem. 1984 May 15;141(1):149-56 PMID: 6373255
  24. Calcium-mediated inhibition of glucuronide production by epinephrine in the perfused rat liver.
    J Biol Chem. 1984 Jun 25;259(12):7705-11 PMID: 6330073
  25. Assembly of asparagine-linked oligosaccharides.
    Annu Rev Biochem. 1985;54:631-64 PMID: 3896128
  26. Egasyn, a protein which determines the subcellular distribution of beta-glucuronidase, has esterase activity.
    J Biol Chem. 1985 Dec 15;260(29):15802-8 PMID: 4066695
  27. Purification and characterization of bovine liver beta-glucuronidase.
    Arch Biochem Biophys. 1966 Sep 26;116(1):193-206 PMID: 6006802
  28. Turnover of constituents of the endoplasmic reticulum membranes of rat hepatocytes.
    J Biol Chem. 1967 May 25;242(10):2389-96 PMID: 4381697
  29. Dual localization of beta-glucuronidase in endoplasmic reticulum and in lysosomes.
    Nature. 1967 Feb 4;213(5075):457-60 PMID: 4166303
  30. Separate genes determining the structure and intracellular location of hepatic glucuronidase.
    Proc Natl Acad Sci U S A. 1967 Sep;58(3):938-45 PMID: 5233850
  31. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  32. Acetanilide-hydrolyzing esterase of rat liver microsomes. I. Solubilization, purification, and intramicrosomal localization.
    J Biochem. 1972 Nov;72(5):1245-56 PMID: 4346266
  33. Biochemical and genetic evidence for a macromolecular -glucuronidase complex in microsomal membranes.
    J Mol Biol. 1973 Jul 5;77(3):371-89 PMID: 4727071
  34. Analytical study of microsomes and isolated subcellular membranes from rat liver. 3. Subfractionation of the microsomal fraction by isopycnic and differential centrifugation in density gradients.
    J Cell Biol. 1974 Apr;61(1):213-31 PMID: 4150490
  35. Posttranslational association of immunoglobulin heavy chain binding protein with nascent heavy chains in nonsecreting and secreting hybridomas.
    J Cell Biol. 1986 May;102(5):1558-66 PMID: 3084497
  36. Identity of esterase-22 and egasyn, the protein which complexes with microsomal beta-glucuronidase.
    Biochem Genet. 1986 Apr;24(3-4):229-43 PMID: 3729927
  37. The egasyn gene affects the processing of oligosaccharides of lysosomal beta-glucuronidase in liver.
    Biochem J. 1986 Dec 1;240(2):445-54 PMID: 3101673
  38. A C-terminal signal prevents secretion of luminal ER proteins.
    Cell. 1987 Mar 13;48(5):899-907 PMID: 3545499
  39. Protein kinase C in fibroblasts. Characteristics of its intracellular location during growth and after exposure to phorbol esters and other mitogens.
    J Biol Chem. 1987 Feb 15;262(5):2234-43 PMID: 3818594
  40. An accessory protein identical to mouse egasyn is complexed with rat microsomal beta-glucuronidase and is identical to rat esterase-3.
    J Biol Chem. 1987 May 25;262(15):7248-53 PMID: 3294829
  41. Involvement of the esterase active site of egasyn in compartmentalization of beta-glucuronidase within the endoplasmic reticulum.
    Cell. 1987 Jul 17;50(2):301-10 PMID: 3594574
  42. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  43. Multiple forms of beta-glucuronidase in rat liver lysosomes and microsomes.
    Arch Biochem Biophys. 1975 Jan;166(1):258-72 PMID: 1122137
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.105.4.1571
Published
1987-10-00
Pages
1571-8
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2114662
Subset
IM
Grants
NIGMS NIH HHS · GM-19521 · United States
NIGMS NIH HHS · GM-33559 · United States
External Links
PubMed source DOI: 10.1083/jcb.105.4.1571 PMC Full Text View journal details
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