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Expression and localization of PCSK9 in rat hepatic cells

Grozdanov, Petar N, Petkov, Petko M, Karagyozov, L K, Dabeva, Mariana D·2006·Biochemistry and Cell Biology
DOI 10.1139/o05-155
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Abstract

Proprotein convertase subtilisin/kexin type 9 (PCSK9), recently cloned in several laboratories, including ours, causes a third form of autosomal dominant hypercholesterolemia. Its mechanism of action remains unclear. We studied the expression and subcellular localization of PCSK9 in fetal and adult rat tissues associated with cholesterol homeostasis using quantitative reverse transcriptase - PCR, Western blot analysis, subcellular fractionation, and confocal immuno fluorescent microscopy. PCSK9 mRNA is most abundant in yolk sac and fetal liver, but the highest expression of the protein was found in differentiated hepa toma FAO-1 cell line, which also shows the highest expression of LDLR. In FAO-1 cells PCSK9 expression is down regulated by cholesterol and 25-hydroxycholesterol and upregulated in the absence of sterols following the same pattern of expression as HMG-CoA reductase, synthase, and LDLR. Subcellular fractionation, combined with Western blotting, showed that PCSK9 is localized in the ER and intermediate vesicular compartment of the cell but not in Golgi cisternae. The mature enzyme is secreted from the liver and hepatoma cells. Double labeling with antibodies to PCSK9 and LDLR or clathrin revealed some colocalization of PCSK9 with clathrin-coated vesicles and LDLR. In conclusion, our results show that PCSK9 is processed in the ER, and the mature convertase is secreted in the plasma.Key words: PCSK9 expression, PCSK9 localization, hepatic cells.

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  • PCSK9 is synthesized as an approximately 75 kDa proprotein (proPCSK9, P-form), and its N-terminal prodomain is removed by autocatalysis generating an approximately 65 kDa C-form that is secreted.

    “PCSK9 is synthesized as an approximately 75 kDa proprotein (proPCSK9, P-form); the N-terminal prodomain is then removed by autocatalysis (Seidah et al, 2003;Benjannet et al, 2004) generating an approximately 65 kDa form (C-form) that is secreted into the extracellular milieu (Grozdanov et al, 2006).”

    From PCSK9 is required for the disposal of non-acetylated intermediates of the nascent membrane protein BACE1 (2008) · cites this paper as Grozdanov et al, 2006

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Metadata

Authors
Grozdanov, P. N., Petkov, P. M., Karagyozov, L. K., & Dabeva, M. D.
Venue
Biochemistry and Cell Biology
Publisher
Canadian Science Publishing
Published
2006-02-01
Volume
84
Issue
1
Pages
80-92
ISSN
  • 0829-8211
  • 1208-6002
DOI
10.1139/o05-155
Cited by
25
References
0
Language
en
Licence
www.nrcresearchpress.com/page/about/CorporateTextAndDataMining (tdm)

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