Molecular Cloning of Human Phosphomevalonate Kinase and Identification of a Consensus Peroxisomal Targeting Sequence

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 (Mr = 21,000-22,500). Further verification of the clones wa...

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Bibliographic Details
Published in:The Journal of biological chemistry 1996-07, Vol.271 (29), p.17330-17334
Main Authors: Chambliss, Ken L., Slaughter, Clive A., Schreiner, Rupert, Hoffmann, Georg F., Gibson, K. Michael
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Brain - enzymology
Cloning, Molecular
Consensus Sequence
DNA Primers
DNA Probes
Female
Humans
Kidney - enzymology
Kinetics
Liver - enzymology
Lymphocytes - enzymology
Molecular Sequence Data
Muscle, Skeletal - enzymology
Myocardium - enzymology
Pancreas - enzymology
Phosphotransferases (Phosphate Group Acceptor) - biosynthesis
Phosphotransferases (Phosphate Group Acceptor) - chemistry
Phosphotransferases (Phosphate Group Acceptor) - metabolism
Placenta - enzymology
Polymerase Chain Reaction
Pregnancy
Recombinant Proteins - biosynthesis
Recombinant Proteins - chemistry
Recombinant Proteins - metabolism
Sequence Homology, Amino Acid
Sequence Homology, Nucleic Acid
Swine
Transcription, Genetic
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Summary: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 (Mr = 21,000-22,500). 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.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.271.29.17330