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PMID: 2508624 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Distribution of G-proteins in rat liver plasma-membrane domains and endocytic pathways.

The Biochemical journal ·Vol. 261 ·No. 3 ·1989-08-01 ·Pages 905-12

Ali N, Milligan G, Evans WH

Abstract

1. The distribution of the alpha- and beta-subunits of nucleotide-binding G-proteins among rat liver sinusoidal, lateral and canalicular plasma membranes, endosomes, Golgi membranes and lysosomes was investigated. 2. Pertussis-toxin-catalysed ADP-ribosylation identified a 41 kDa inhibitory alpha-subunit in all liver plasma-membrane functional domains as well as in endosomes. An antibody to a synthetic peptide corresponding to a C-terminal sequence of the inhibitory alpha-subunit also identified the 41 kDa polypeptide in all plasma-membrane domains, in 'early' and 'late' endosomes and in Golgi membranes; this polypeptide was not detected in lysosomes. The antibody-binding studies showed that bile-canalicular plasma membranes had the highest content of the inhibitory alpha-subunit. 3. Immunofluorescent microscopy confirmed the presence of the inhibitory alpha-subunit in all regions of the hepatocyte's cell surface. 4. An antibody recognizing the beta-subunit showed that a 36 kDa polypeptide was present in all plasma membranes and in 'early' and 'late' endosomes; it was not detected in lysosomes. The relative distribution among the fractions of this polypeptide was similar to the distribution of the inhibitory alpha-subunit. 5. The presence of high levels of the G-protein inhibitory alpha-subunit in bile-canalicular plasma membranes was confirmed by demonstration of its co-fractionation with marker enzymes in Nycodenz gradients and by free-flow electrophoresis. The significance of this location is discussed.

MeSH Terms
Animals Bile Canaliculi/analysis Blotting, Western Cell Membrane/analysis Endocytosis GTP-Binding Proteins/analysis Liver/analysis,ultrastructure Lysosomes/analysis Male Organelles/analysis Rats Rats, Inbred Strains
Chemicals
GTP-Binding Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ali N
National Institute for Medical Research, Mill Hill, London, U.K.
Milligan G
Evans W H
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42 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2610905
Published
1989-08-01
Pages
905-12
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1138915
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1042/bj2610905 PMC Full Text View journal details
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