Molecular cloning of human phosphomevalonate kinase and identification of a consensus peroxisomal targeting sequence

Ken L. Chambliss, Clive A. Slaughter, Rupert Schreiner, Georg F. Hoffmann, K. Michael Gibson

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Two overlapping cDNAs which encode human liver phosphomevalonate kinase (PMKase) were isolated. The human PMKase cDNAs predict a 191-amino acid protein with a molecular weight of 21,862, consistent with previous reports for mammalian PMKase (M(r) = 21,00022,500). Further verification of the clones was obtained by expression of PMKase activity in bacteria using a composite 1024-base pair cDNA clone. Northern blot analysis of several human tissues revealed a doublet of transcripts at approximately 1 kilobase (kb) in heart, liver, skeletal muscle, kidney, and pancreas and lower but detectable transcript levels in brain, placenta, and lung. Analysis of transcripts from human lymphoblasts subcultured in lipid-depleted sera (LDS) and LDS supplemented with lovastatin indicated that PMKase gene expression is subject to regulation by sterol at the level of transcription. Southern blotting indicated that PMKase is a single copy gene covering less than 15 kb in the human genome. The human PMKase amino acid sequence contains a consensus peroxisomal targeting sequence (PTS-1), Ser-Arg-Leu, at the C terminus of the protein. This is the first report of a cholesterol biosynthetic protein which contains a consensus PTS-1, providing further evidence for the concept that early cholesterol and nonsterol isoprenoid biosynthesis may occur in the peroxisome.

Original languageEnglish (US)
Pages (from-to)17330-17334
Number of pages5
JournalJournal of Biological Chemistry
Volume271
Issue number29
DOIs
StatePublished - Aug 5 1996
Externally publishedYes

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Cloning
Molecular Cloning
Complementary DNA
Liver
Clone Cells
Genes
Cholesterol
Lipids
Amino Acids
Lovastatin
Peroxisomes
Biosynthesis
Terpenes
Sterols
Human Genome
Transcription
Southern Blotting
Protein C
phosphomevalonate kinase
Serum

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular cloning of human phosphomevalonate kinase and identification of a consensus peroxisomal targeting sequence. / Chambliss, Ken L.; Slaughter, Clive A.; Schreiner, Rupert; Hoffmann, Georg F.; Michael Gibson, K.

In: Journal of Biological Chemistry, Vol. 271, No. 29, 05.08.1996, p. 17330-17334.

Research output: Contribution to journalArticle

Chambliss, Ken L. ; Slaughter, Clive A. ; Schreiner, Rupert ; Hoffmann, Georg F. ; Michael Gibson, K. / Molecular cloning of human phosphomevalonate kinase and identification of a consensus peroxisomal targeting sequence. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 29. pp. 17330-17334.
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