Abstract
In this paper we describe isolation and molecular characterization of human dihydroxyacetonephosphate acyltransferase (DAP-AT). The enzyme was extracted from rabbit Harderian gland peroxisomes and isolated as a trimeric complex by sucrose density gradient centrifugation. From peptide sequences matching EST-clones were obtained which allowed cloning and sequencing of the cDNA from a human cDNA library. The nucleotide-derived amino acid sequence revealed a protein consisting of 680 amino acid residues of molecular mass 77187 containing a C-terminal type 1 peroxisomal targeting signal. Monospecific antibodies raised against this polypeptide efficiently immunoprecipitated DAP-AT activity from solubilized peroxisomal preparations, thus demonstrating that the cloned cDNA codes for DAP-AT.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acyltransferases / analysis
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Acyltransferases / chemistry*
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Acyltransferases / isolation & purification
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Acyltransferases / metabolism
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Amino Acid Sequence
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Animals
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Base Sequence
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Brain / enzymology
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Centrifugation, Density Gradient
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Harderian Gland / enzymology
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Humans
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Microbodies / enzymology*
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Microscopy, Immunoelectron
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Molecular Sequence Data
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Peptide Fragments / chemistry
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Peptide Fragments / immunology
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Peptide Fragments / isolation & purification
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Phospholipid Ethers / metabolism
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Precipitin Tests
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Protein Sorting Signals / chemistry
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Rabbits
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Sequence Analysis, DNA
Substances
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Peptide Fragments
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Phospholipid Ethers
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Protein Sorting Signals
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Acyltransferases
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glycerone-phosphate O-acyltransferase