Mutagenesis of the human apolipoprotein B gene in a yeast artificial chromosome reveals the site of attachment for apolipoprotein(a)

Sally P A Mccormick, Jennifer K. Ng, Stacy Taylor, Laura M. Flynn, Robert E Hammer, Stephen G. Young

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Lipoprotein(a) [Lp(a)] is a lipoprotein formed by the disulfide linkage of apolipoprotein (apo) B100 of a low density lipoprotein particle to apolipoprotein(a). Prior studies have suggested that one of the C-terminal Cys residues of apo-B100 is involved in the disulfide linkage of apo-B100 to apo(a). To identify the apo-B100 Cys residue involved in the formation of Lp(a), we constructed a yeast artificial chromosome (YAC) spanning the human apo-B gene and used gene-targeting techniques to change Cys-4326 to Gly. The mutated YAC DNA was used to generate transgenic mice expressing the mutant human apo-B100 (Cys4326Gly). Unlike the wild-type human apo-B100, the mutant human apo-B100 completely lacked the ability to bind to apo(a) and form Lp(a). This study demonstrates that apo-B100 Cys-4326 is required for the assembly of Lp(a) and shows that gene targeting in YACs, followed by the generation of transgenic mice, is a useful approach for analyzing the structure of large proteins coded for by large genes.

Original languageEnglish (US)
Pages (from-to)10147-10151
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number22
DOIs
StatePublished - Oct 24 1995

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Yeast Artificial Chromosomes
Apoprotein(a)
Apolipoproteins
Apolipoproteins B
Mutagenesis
Lipoprotein(a)
Genes
Gene Targeting
Disulfides
Transgenic Mice
LDL Lipoproteins
Lipoproteins
DNA

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Mutagenesis of the human apolipoprotein B gene in a yeast artificial chromosome reveals the site of attachment for apolipoprotein(a). / Mccormick, Sally P A; Ng, Jennifer K.; Taylor, Stacy; Flynn, Laura M.; Hammer, Robert E; Young, Stephen G.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 22, 24.10.1995, p. 10147-10151.

Research output: Contribution to journalArticle

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