Lecithin:cholesterol acyltransferase deficiency: Identification of two defective alleles in fibroblast cDNA

M. Miller, K. Zeller, P. C. Kwiterovich, J. J. Albers, G. Feulner

Research output: Contribution to journalArticlepeer-review

Abstract

Previous mutations associated with lecithin:cholesterol acyltransferase (LCAT) deficiency have been identified using genomic DNA. To facilitate mutation analysis, we used cDNA front cultured fibroblasts which were shown to express LCAT mRNA. Using reverse-transcriptase PCR, LCAT cDNA was obtained thorn a 13-year-old boy with complete LCAT deficiency, characterized by low HDL-C (3 mg/dl), nondetectable initial cholesterol esterification rate, LCAT activity, and minimal LCAT mass (0.16 vs. 5-7.5 μg/ml). Sequencing of LCAT cDNA clones identified two mutations. A novel frame-shift mutation caused by deletion of cytosine at the third nucleotide position of amino acid 168 (exon 5) predicts a disrupted protein catalytic site by converting Ser181→Ala and creates a Pvu-II restriction site prior to premature truncation at amino acid 238. A C→T transition results in a substitution of methionine for threonine at amino acid position 321 and creates an Nla-III restriction site on the maternal allele. Expression studies of mutant LCAT cDNA confirmed the virtual absence of LCAT activity in transfected COS-1 cells. The molecular defect in a young male with complete LCAT deficiency has been identified using fibroblast cDNA.

Original languageEnglish (US)
Pages (from-to)931-938
Number of pages8
JournalJournal of Lipid Research
Volume36
Issue number5
StatePublished - 1995

Keywords

  • HDL-C
  • LCAT
  • complementary DNA
  • fibroblasts
  • genetic disease

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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