On the spatial arrangement of proteins in microsomal membranes from rat liver

G. Kreibich, Ann Louise Hubbard, D. D. Sabatini

Research output: Contribution to journalArticle

Abstract

Rat liver rough microsomes were labeled enzymatically with 125I using lactoperoxidase and glucose oxidase. In intact microsomes only proteins exposed on the outside face of the microsomal membrane were iodinated. Low concentrations of detergent (0.049% deoxycholate) were used to allow entrance of the iodination system into the vesicles without disassembling the membranes. This led to iodination of the soluble content proteins and to an increased labeling of the membrane proteins. The distribution of radioactivity in microsomal proteins was analyzed after separation by sodium dodecyl sulfate acrylamide gel electrophoresis. Most membrane proteins were labeled when intact microsomes were iodinated. No major membrane proteins were exclusively labeled in the presence of low detergent concentrations or after complete membrane disassembly. Therefore it is unlikely that there are major membrane proteins, other than glycoproteins, present only on the inner membrane face or completely embedded within the microsomal membrane. Microsomal proteins were also labeled by incubating rough microsomes with [3H] NaBH4 after reaction with pyridoxal phosphate. Microsomal membranes were permeable to these small molecular weight reagents as shown by the fact that proteins in the vesicular cavity as well as membrane proteins were labeled with this system.

Original languageEnglish (US)
Pages (from-to)616-627
Number of pages12
JournalJournal of Cell Biology
Volume60
Issue number3
StatePublished - 1974
Externally publishedYes

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Membrane Proteins
Membranes
Liver
Microsomes
Proteins
Halogenation
Detergents
Lactoperoxidase
Glucose Oxidase
Pyridoxal Phosphate
Deoxycholic Acid
Acrylamide
Liver Microsomes
Sodium Dodecyl Sulfate
Radioactivity
Electrophoresis
Glycoproteins
Molecular Weight
Gels

ASJC Scopus subject areas

  • Cell Biology

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On the spatial arrangement of proteins in microsomal membranes from rat liver. / Kreibich, G.; Hubbard, Ann Louise; Sabatini, D. D.

In: Journal of Cell Biology, Vol. 60, No. 3, 1974, p. 616-627.

Research output: Contribution to journalArticle

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