Influence of allelic variation on apolipoprotein(a) folding in the endoplasmic reticulum

Ann L. White, Bernadette Guerra, Robert E. Lanford

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Plasma levels of lipoprotein(a) (Lp(a)) vary over 1000-fold between individuals and are determined by the gene for its unique apolipoprotein, apo(a), which has greater than 100 alleles. Using primary baboon hepatocyte cultures, we previously demonstrated that differences in the ability of apo(a) allelic variants to escape the endoplasmic reticulum (ER) are a major determinant of Lp(a) production rate. To examine the reason for these differences, the folding of newly synthesized apo(a) was analyzed in pulse- chase experiments. Samples were harvested in the presence of N- ethylmaleimide to preserve disulfide-bonded folding intermediates, and apo(a) was analyzed by immunoprecipitation and SDS-polyacrylamide gel electrophoresis. Apo(a) required a prolonged period (30-60 min) to reach its fully oxidized form. Multiple folding intermediates were resolved, including a disulfide-linked, apo(a)-containing complex. Unexpectedly, all allelic variants examined showed similar patterns and kinetics of folding. Even 'null' apo(a) proteins, which are unable to exit the ER, appeared to fold normally. The ER glucosidase inhibitor, castanospermine, prevented apo(a) secretion, but did not inhibit folding. This suggests that an event which is dependent on trimming of N-linked glucoses, and which occurs after the folding events detectable in our assay, is required for apo(a) secretion. Differences in the ability to undergo this event may explain the variable efficiency with which apo(a) allelic variants exit the ER.

Original languageEnglish (US)
Pages (from-to)5048-5055
Number of pages8
JournalJournal of Biological Chemistry
Volume272
Issue number8
DOIs
StatePublished - Feb 21 1997
Externally publishedYes

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Apoprotein(a)
Lipoprotein(a)
Endoplasmic Reticulum
Disulfides
Glucosidases
Ethylmaleimide
Trimming
Apolipoproteins
Electrophoresis
Assays
Genes
Plasmas
Glucose
Kinetics
Papio
Immunoprecipitation
Polyacrylamide Gel Electrophoresis
Hepatocytes
Proteins
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Influence of allelic variation on apolipoprotein(a) folding in the endoplasmic reticulum. / White, Ann L.; Guerra, Bernadette; Lanford, Robert E.

In: Journal of Biological Chemistry, Vol. 272, No. 8, 21.02.1997, p. 5048-5055.

Research output: Contribution to journalArticle

White, Ann L. ; Guerra, Bernadette ; Lanford, Robert E. / Influence of allelic variation on apolipoprotein(a) folding in the endoplasmic reticulum. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 8. pp. 5048-5055.
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