Document Detail


Characterization of recombinant human apoB-48-containing lipoproteins in rat hepatoma McA-RH7777 cells transfected with apoB-48 cDNA. Overexpression of apoB-48 decreases synthesis of endogenous apoB-100.
MedLine Citation:
PMID:  7749860     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We studied the effect of overexpression of apolipoprotein (apo) B-48 on the synthesis and secretion of endogenous apoB-100 in rat hepatoma McA-RH7777 cell lines stably transfected with human apoB-48 cDNA under the control of the cytomegalovirus promoter. Three cell lines that secrete 40 to 60 ng human apoB.mg cell protein-1.h-1 were used. The recombinant human apoB-48 exhibited physicochemical characteristics (buoyant density, 1.06 to 1.21 g/mL; beta-electrophoretic mobility and diameters, 16 to 20 nm) indistinguishable from those of endogenous rat apoB-48. Overexpression of the recombinant human apoB-48 resulted in a 50% decrease in the secretion of endogenous apoB-100 but did not affect the secretion of apoE or apoA-I. Several possible mechanisms for the decreased secretion of apoB-100 were evaluated. First, recruitment of lipids into lipoproteins was shown to be unaffected since no major changes in the physicochemical properties of apoB-100-containing lipoproteins were observed. Second, the intracellular degradation of apoB-100 was not altered as the intracellular retention half-time and secretion efficiency remained unaffected by apoB-48 overexpression. Third, the posttranslational regulatory mechanisms for apoB-100 remained normal, as demonstrated by a twofold increase in apoB-100 secretion after supplementation with oleic acid. Unexpectedly, a 35% to 50% decrease in the steady-state synthesis of endogenous apoB-100 was observed in apoB-48-transfected cells compared with control cells. These data suggested that decreased secretion of apoB-100 was secondary to decreased synthesis. The decreased apoB-100 synthesis was not due to decreased steady-state levels of rat apoB-100 mRNA. These results suggest that overexpression of recombinant human apoB-48 may interfere with posttranscriptional events, possibly at the translation-translocation level, and decrease translational yield of apoB-100. These posttranscriptional events prior to the complete synthesis of the apoB-100 polypeptide can be important in the control of apoB-100 secretion.
Authors:
M M Hussain; Y Zhao; R K Kancha; B D Blackhart; Z Yao
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Arteriosclerosis, thrombosis, and vascular biology     Volume:  15     ISSN:  1079-5642     ISO Abbreviation:  Arterioscler. Thromb. Vasc. Biol.     Publication Date:  1995 Apr 
Date Detail:
Created Date:  1995-06-22     Completed Date:  1995-06-22     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9505803     Medline TA:  Arterioscler Thromb Vasc Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  485-94     Citation Subset:  IM    
Affiliation:
Department of Pathology, Medical College of Pennsylvania, Philadelphia 19129, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Apolipoprotein B-100
Apolipoprotein B-48
Apolipoproteins B / biosynthesis,  genetics*,  metabolism
Base Sequence
Carcinoma, Hepatocellular / genetics,  pathology
DNA, Complementary / genetics
Humans
Lipoproteins / chemistry,  genetics*
Molecular Sequence Data
Rats
Recombinant Proteins / biosynthesis,  genetics,  metabolism
Transfection
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Apolipoprotein B-100; 0/Apolipoprotein B-48; 0/Apolipoproteins B; 0/DNA, Complementary; 0/Lipoproteins; 0/Recombinant Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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