Biochemical fractionation and characterization of proteins from Golgi-enriched membranes

Subramaniam, V. N., Bin Mohd Yusoff, A. R., Wong, S. H., Lim, G. B., Chew, M. and Hong, W. J. (1992) Biochemical fractionation and characterization of proteins from Golgi-enriched membranes. Journal of Biological Chemistry, 267 17: 12016-12021.

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Author Subramaniam, V. N.
Bin Mohd Yusoff, A. R.
Wong, S. H.
Lim, G. B.
Chew, M.
Hong, W. J.
Title Biochemical fractionation and characterization of proteins from Golgi-enriched membranes
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
1083-351X
Publication date 1992-06-15
Sub-type Article (original research)
Open Access Status File (Publisher version)
Volume 267
Issue 17
Start page 12016
End page 12021
Total pages 6
Place of publication Bethesda, MD, United States
Publisher American Society for Biochemistry and Molecular Biology
Language eng
Abstract Fractions enriched in Golgi membranes were prepared from rat liver by sucrose gradient ultracentrifugation. These enriched membranes were further subfractionated on the basis of their solubilities in EGTA, 150 mM sodium carbonate, pH 11.5, sodium deoxycholate, Triton X-100, or sodium dodecyl sulfate. This led to isolation of peripheral, luminal, and integral membrane proteins of the Golgi-enriched membranes. Luminal and membrane proteins were further purified by wheat germ agglutinin and concanavalin A lectin affinity chromatographies. Some proteins from these lectin columns were resolved by preparative gel electrophoresis and microsequenced. Subsequently, antibodies were produced for two proteins by immunization of either mice or rabbits. Immunofluorescence microscopy suggests that these proteins are confined to Golgi apparatus-like structures. The protocol described is well suited for the study of organelle structure and function.
Keyword Rat liver
Endoplasmic reticulum
Binding protein
Cdna Cloning
Apparatus
Identification
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Medicine Publications
 
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