Home LiteratureArticle Details
PMID: 6470038 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Biosynthesis and processing of ribophorins in the endoplasmic reticulum.

The Journal of cell biology ·Vol. 99 ·No. 3 ·1984-09-00 ·Pages 1076-82

Rosenfeld MG, Marcantonio EE, Hakimi J, Ort VM, Atkinson PH, Sabatini D, Kreibich G

Abstract

Ribophorins are two transmembrane glycoproteins characteristic of the rough endoplasmic reticulum, which are thought to be involved in the binding of ribosomes. Their biosynthesis was studied in vivo using lines of cultured rat hepatocytes (clone 9) and pituitary cells (GH 3.1) and in cell-free synthesis experiments. In vitro translation of mRNA extracted from free and bound polysomes of clone 9 cells demonstrated that ribophorins are made exclusively on bound polysomes. The primary translation products of ribophorin messengers obtained from cultured hepatocytes or from regenerating livers co-migrated with the respective mature proteins, but had slightly higher apparent molecular weights (2,000) than the unglycosylated forms immunoprecipitated from cells treated with tunicamycin. This indicates that ribophorins, in contrast to all other endoplasmic reticulum membrane proteins previously studied, contain transient amino-terminal insertion signals which are removed co-translationally. Kinetic and pulse-chase experiments with [35S]methionine and [3H]mannose demonstrated that ribophorins are not subjected to electrophoretically detectable posttranslational modifications, such as proteolytic cleavage or trimming and terminal glycosylation of oligosaccharide side chain(s). Direct analysis of the oligosaccharides of ribophorin l showed that they do not contain the terminal sugars characteristic of complex oligosaccharides and that they range in composition from Man8GlcNAc to Man5GlcNAc. These findings, as well as the observation that the mature proteins are sensitive to endoglycosidase H and insensitive to endoglycosidase D, are consistent with the notion that the biosynthetic pathway of the ribophorins does not require a stage of passage through the Golgi apparatus.

MeSH Terms
Animals Cell Line Endoplasmic Reticulum/metabolism Intracellular Membranes/metabolism Liver Membrane Proteins/biosynthesis,genetics Microsomes/metabolism Oligosaccharides/analysis Plants/metabolism Protein Biosynthesis Protein Processing, Post-Translational RNA, Messenger/genetics Rats Ribosomes/metabolism Triticum/metabolism
Chemicals
Membrane Proteins Oligosaccharides RNA, Messenger ribophorin
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Rosenfeld M G
Marcantonio E E
Hakimi J
Ort V M
Atkinson P H
Sabatini D
Kreibich G
References (54)
54 references, click to expand
  1. Cell-surface glycopeptides: growth-dependent changes in the carbohydrate-peptide linkage region.
    J Mol Biol. 1973 Nov 15;80(4):781-99 PMID: 4773030
  2. Rotation-mediated histogenetic aggregation of dissociated cells. A quantifiable approach to cell interactions in vitro.
    Exp Cell Res. 1961 Jan;22:455-75 PMID: 13773012
  3. A procedure for the quantitative recovery of homogeneous populations of undegraded free and bound polysomes from rat liver.
    Biochemistry. 1976 Apr 20;15(8):1704-12 PMID: 1268192
  4. Purification and characterization of alpha-D-mannosidase from rat liver golgi membranes.
    J Biol Chem. 1977 May 25;252(10):3227-33 PMID: 863880
  5. Rat insulin genes: construction of plasmids containing the coding sequences.
    Science. 1977 Jun 17;196(4296):1313-9 PMID: 325648
  6. Efficient cleavage and segregation of nascent presecretory proteins in a reticulocyte lysate supplemented with microsomal membranes.
    J Biol Chem. 1978 Jun 10;253(11):3753-6 PMID: 649601
  7. Proteins of rough microsomal membranes related to ribosome binding. I. Identification of ribophorins I and II, membrane proteins characteristics of rough microsomes.
    J Cell Biol. 1978 May;77(2):464-87 PMID: 649658
  8. Proteins of rough microsomal membranes related to ribosome binding. II. Cross-linking of bound ribosomes to specific membrane proteins exposed at the binding sites.
    J Cell Biol. 1978 May;77(2):488-506 PMID: 418074
  9. Biochemical studies of rat liver Golgi apparatus. I. Isolation and preliminary characterization.
    J Biochem. 1978 Apr;83(4):909-23 PMID: 96108
  10. Biochemical studies on rat liver Golgi apparatus. II. Further characterization of isolated Golgi fraction.
    J Biochem. 1978 Apr;83(4):925-34 PMID: 207681
  11. Biochemical studies on rat liver Golgi apparatus. III. Subfractionation of fragmented Golgi apparatus by counter-current distribution.
    J Biochem. 1978 Apr;83(4):935-42 PMID: 96109
  12. The synthesis of complex-type oligosaccharides. III. Identification of an alpha-D-mannosidase activity involved in a late stage of processing of complex-type oligosaccharides.
    J Biol Chem. 1978 Nov 10;253(21):7779-86 PMID: 212436
  13. Distribution of phospholipid biosynthetic enzymes among cell components of rat liver.
    J Biol Chem. 1978 Nov 10;253(21):7960-71 PMID: 212440
  14. Spatial orientation of glycoproteins in membranes of rat liver rough microsomes. II. Transmembrane disposition and characterization of glycoproteins.
    J Cell Biol. 1978 Sep;78(3):894-909 PMID: 701364
  15. Endoplasmic reticulum marker enzymes in Golgi fractions--what does this mean?
    J Cell Biol. 1978 Nov;79(2 Pt 1):581-9 PMID: 214450
  16. Presence of NADPH-cytochrome P-450 reductase in rat liver Golgi membranes. Evidence obtained by immunoadsorption method.
    J Cell Biol. 1978 Nov;79(2 Pt 1):590-7 PMID: 214451
  17. Characterization of the ribosomal binding site in rat liver rough microsomes: ribophorins I and II, two integral membrane proteins related to ribosome binding.
    J Supramol Struct. 1978;8(3):279-302 PMID: 723266
  18. In vitro synthesis of the Ca2+ transport ATPase by ribosomes bound to sarcoplasmic reticulum membranes.
    Proc Natl Acad Sci U S A. 1979 Mar;76(3):1241-5 PMID: 155818
  19. Messenger RNA of opsin from bovine retina: isolation and partial sequence of the in vitro translation product.
    Proc Natl Acad Sci U S A. 1979 Jun;76(6):2654-8 PMID: 288054
  20. Purification and characterization of a rat liver Golgi alpha-mannosidase capable of processing asparagine-linked oligosaccharides.
    J Biol Chem. 1979 Nov 25;254(22):11655-63 PMID: 500665
  21. Biological detection of specific mRNA molecules by microinjection.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4503-6 PMID: 291982
  22. Identification of ribophorins in rough microsomal membranes from different organs of several species.
    Eur J Biochem. 1982 May;124(1):217-22 PMID: 7084228
  23. Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation.
    J Biol Chem. 1983 Mar 10;258(5):3159-65 PMID: 6402509
  24. O-linked oligosaccharides are acquired by herpes simplex virus glycoproteins in the Golgi apparatus.
    Cell. 1983 Mar;32(3):987-97 PMID: 6299584
  25. In vitro synthesis and membrane insertion of bovine MP26, an integral protein from lens fiber plasma membrane.
    J Cell Biol. 1983 Mar;96(3):633-8 PMID: 6833374
  26. Evidence for an alpha-mannosidase in endoplasmic reticulum of rat liver.
    J Biol Chem. 1983 Jul 10;258(13):7907-10 PMID: 6863271
  27. Hepatoma secretory proteins migrate from rough endoplasmic reticulum to Golgi at characteristic rates.
    Nature. 1983 Jul 7-13;304(5921):80-3 PMID: 6866094
  28. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose.
    Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408-12 PMID: 4504350
  29. Characterization of poly(A+)RNA in free messenger ribonucleoprotein and polysomes of mouse Taper ascites cells.
    J Mol Biol. 1979 Aug 25;132(4):679-94 PMID: 529278
  30. Synthesis and insertion of cytochrome P-450 into endoplasmic reticulum membranes.
    Proc Natl Acad Sci U S A. 1980 Feb;77(2):965-9 PMID: 6767247
  31. Localization and biosynthesis of NADH-cytochrome b5 reductase, an integral membrane protein, in rat liver cells. I. Distribution of the enzyme activity in microsomes, mitochondria, and golgi complex.
    J Cell Biol. 1980 Jun;85(3):501-15 PMID: 7391131
  32. Biosynthesis of erythrocyte membrane protein band 3 in DMSO-induced Friend erythroleukemia cells.
    J Cell Physiol. 1980 Aug;104(2):261-8 PMID: 6931832
  33. Growth-dependent alterations in oligomannosyl glycopeptides expressed in Sindbis virus glycoproteins.
    Biochemistry. 1980 Nov 25;19(24):5619-24 PMID: 7459334
  34. Characterization of the oligosaccharides of liver Z variant alpha 1-antitrypsin.
    Can J Biochem. 1980 Aug;58(8):644-8 PMID: 6970063
  35. Structural gene products of the Ah complex. Increases in large mRNA from mouse liver associated with cytochrome P1-450 induction by 3-methylcholanthrene.
    J Biol Chem. 1981 Mar 25;256(6):3085-91 PMID: 7204392
  36. The golgi apparatus: two organelles in tandem.
    Science. 1981 Sep 11;213(4513):1212-9 PMID: 7268428
  37. Synthesis and processing of asparagine-linked oligosaccharides.
    Annu Rev Biochem. 1981;50:555-83 PMID: 7023366
  38. Processing of the carbohydrate units of thyroglobulin.
    J Biol Chem. 1981 Oct 10;256(19):10161-8 PMID: 7275973
  39. Co-translational membrane integration of calcium pump protein without signal sequence cleavage.
    Nature. 1981 Jul 2;292(5818):87-8 PMID: 6456415
  40. Effect of energy depletion on the glycosylation of a viral glycoprotein.
    J Biol Chem. 1981 Nov 10;256(21):11191-8 PMID: 7287762
  41. Mechanisms for the incorporation of proteins in membranes and organelles.
    J Cell Biol. 1982 Jan;92(1):1-22 PMID: 7035466
  42. Studies on the biosynthesis of microsomal membrane proteins. Site of synthesis and mode of insertion of cytochrome b5, cytochrome b5 reductase, cytochrome P-450 reductase and epoxide hydrolase.
    Eur J Biochem. 1982 Feb;122(2):393-402 PMID: 6800789
  43. alpha-D-Mannosidases of rat liver Golgi membranes. Mannosidase II is the GlcNAcMAN5-cleaving enzyme in glycoprotein biosynthesis and mannosidases Ia and IB are the enzymes converting Man9 precursors to Man5 intermediates.
    J Biol Chem. 1982 Apr 10;257(7):3660-8 PMID: 7061502
  44. Vesicular stomatitis virus glycoprotein, albumin, and transferrin are transported to the cell surface via the same Golgi vesicles.
    J Cell Biol. 1983 Dec;97(6):1815-22 PMID: 6315744
  45. 3-Hydroxy-3-methylglutaryl-CoA reductase: a transmembrane glycoprotein of the endoplasmic reticulum with N-linked "high-mannose" oligosaccharides.
    Proc Natl Acad Sci U S A. 1983 Dec;80(23):7165-9 PMID: 6580634
  46. Biosynthesis of N- and O-linked oligosaccharides of the low density lipoprotein receptor.
    J Biol Chem. 1983 Dec 25;258(24):15261-73 PMID: 6317691
  47. Biosynthesis of hepatocyte endoplasmic reticulum proteins.
    Methods Enzymol. 1983;96:530-42 PMID: 6656643
  48. Processing of MOPC 315 immunoglobulin A oligosaccharides: evidence for endoplasmic reticulum and trans Golgi alpha 1,2-mannosidase activity.
    J Cell Biol. 1984 Feb;98(2):407-16 PMID: 6420419
  49. Nucleotide sequence of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase, a glycoprotein of endoplasmic reticulum.
    Nature. 1984 Apr 12-18;308(5960):613-7 PMID: 6546784
  50. Assembly in vitro of a spanning membrane protein of the endoplasmic reticulum: the E1 glycoprotein of coronavirus mouse hepatitis virus A59.
    Proc Natl Acad Sci U S A. 1984 Mar;81(5):1421-5 PMID: 6324191
  51. Biogenesis of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, an integral glycoprotein of the endoplasmic reticulum.
    Proc Natl Acad Sci U S A. 1984 Mar;81(6):1674-8 PMID: 6584901
  52. Viral glycoproteins destined for apical or basolateral plasma membrane domains traverse the same Golgi apparatus during their intracellular transport in doubly infected Madin-Darby canine kidney cells.
    J Cell Biol. 1984 Apr;98(4):1304-19 PMID: 6325468
  53. Co-translational excision of alpha-glucose and alpha-mannose in nascent vesicular stomatitis virus G protein.
    J Cell Biol. 1984 Jun;98(6):2245-9 PMID: 6327729
  54. Tunicamycin--an inhibitor of yeast glycoprotein synthesis.
    Biochem Biophys Res Commun. 1974 May 7;58(1):287-95 PMID: 4598444
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.99.3.1076
Published
1984-09-00
Pages
1076-82
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113407
Subset
IM
Grants
NCI NIH HHS · CA 13402 · United States
NIGMS NIH HHS · GM 20277 · United States
NIGMS NIH HHS · GM 21971 · United States
External Links
PubMed source DOI: 10.1083/jcb.99.3.1076 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2