cDNA cloning of the human tumor motility-stimulating protein, autotaxin, reveals a homology with phosphodiesterases.
A human cDNA clone encoding autotaxin, a tumor cell motility-stimulating protein, reveals that this protein is an ecto/exo-enzyme with significant homology to the plasma cell membrane differentiation antigen PC-1. ATX is a 125-kDa glycoprotein, previously isolated from a human melanoma cell line (A2058), which elicits chemotactic and chemokinetic responses at picomolar to nanomolar concentrations. Affinity-purified antipeptide antibodies to the ATX peptide, ATX-102, were employed to screen an A2058 cDNA expression library made in lambda gt11. The partial cDNA sequence which was obtained was then extended by utilizing reverse transcriptase on total cellular RNA followed by polymerase chain reaction amplification. The isolated cDNA clone contained 3251 base pairs, and the mRNA message size was approximately 3.3 kilobases. The deduced amino acid sequence of autotaxin matched 30 previously sequenced peptides and comprised a protein of 915 amino acids. Data base analysis of the ATX sequence revealed a 45% amino acid identity (including 30 out of 33 cysteines) with PC-1, a pyrophosphatase/type I phosphodiesterase expressed on the surface of activated B cells and plasma cells. ATX, like PC-1, was found to hydrolyze the type I phosphodiesterase substrate p-nitrophenyl thymidine-5'-monophosphate. Autotaxin now defines a novel motility-regulating function for this class of ecto/exo-enzymes.
Murata
J
J
Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Lee
H Y
HY
Clair
T
T
Krutzsch
H C
HC
Arestad
A A
AA
Sobel
M E
ME
Liotta
L A
LA
Stracke
M L
ML
D000595:Amino Acid Sequence; D000906:Antibodies; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D008545:Melanoma; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011755:Pyrophosphatases; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured; D000099189:Lysophospholipase D
D000906:Antibodies; D017931:DNA Primers; D018076:DNA, Complementary; D005956:Glucose-6-Phosphate Isomerase; D006023:Glycoproteins; D009097:Multienzyme Complexes; D010455:Peptides; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases; D000099189:Lysophospholipase D