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PMID: 6304108 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

A domain-specific marker for the hepatocyte plasma membrane: localization of leucine aminopeptidase to the bile canalicular domain.

The Journal of cell biology ·Vol. 96 ·No. 6 ·1983-06-00 ·Pages 1548-58

Roman LM, Hubbard AL

Abstract

Indirect immunofluorescence was used to establish a domain-specific marker for hepatocyte plasma membranes. In frozen sections of fixed rat liver (0.5-4 microns), antibodies directed against rat intestinal leucine aminopeptidase (LAP) recognized an antigen that was restricted to the bile canalicular plasma membrane. Fluorescence was not observed on the sinusoidal or lateral membranes, and intracellular staining was not detected. The liver antigen was identified as LAP, based on its chemical similarity to intestinal LAP. First, immunoprecipitation experiments using trypsin-solubilized intestinal LAP (G-200 fraction, 91% pure) established a correlation between the loss of LAP enzyme activity from the soluble fraction and the appearance in the specific immunoprecipitates of polypeptides migrating on SDS PAGE between 110,000 and 130,000 daltons. The antigen precipitated from a detergent extract of liver plasma membranes had the same electrophoretic mobility. Second, the chymotryptic map of the major band in the liver immunoprecipitate was similar to that of purified intestinal LAP.

MeSH Terms
Acid Anhydride Hydrolases Alkaline Phosphatase/analysis Animals Bile Ducts/cytology Cell Membrane/enzymology Chickens Chymotrypsin/metabolism Fluorescent Antibody Technique Guinea Pigs Immunodiffusion Intestine, Small/cytology Leucyl Aminopeptidase/analysis Liver/cytology Phosphoric Monoester Hydrolases/analysis Rats
Chemicals
Alkaline Phosphatase Phosphoric Monoester Hydrolases Leucyl Aminopeptidase Chymotrypsin Acid Anhydride Hydrolases trimetaphosphatase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Roman L M
Hubbard A L
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41 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.96.6.1548
Published
1983-06-00
Pages
1548-58
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2112471
Subset
IM
Grants
NIGMS NIH HHS · GM29185 · United States
External Links
PubMed source DOI: 10.1083/jcb.96.6.1548 PMC Full Text View journal details
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