Site-directed mutations in the tumor-associated cytokine, autotaxin, eliminate nucleotide phosphodiesterase, lysophospholipase D, and motogenic activities.
The exo-enzyme autotaxin/NPP2 (ATX/NPP2) is a potent stimulator of cell migration, invasion, metastasis, and angiogenesis. Recently, ATX/NPP2 was found to possess lysophospholipase D (lyso-LPD) activity, generating the bioactive mediator lysophosphatidic acid from precursors. In the present study, we used site-directed mutagenesis to delineate the active domain of lysophospholipid catalytic activity and to examine potential overlap with the nucleotide phosphodiesterase domain. We found four amino acid residues obligatory for the phosphodiesterase, lyso-PLD, and migration-stimulating activities of ATX/NPP2, suggesting that 5'-nucleotide phosphodiesterase (PDE) and lyso-PLD share a common reaction mechanism and inviting design of enzymatic inhibitors as therapeutic agents for neoplastic disease.
Laboratory of Pathology, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA. kohe@mail.nih.gov
Eunjin
E
Koh
Timothy
T
Clair
Elisa C
EC
Woodhouse
Elliott
E
Schiffmann
Lance
L
Liotta
Mary
M
Stracke
D000818:Animals; D019556:COS Cells; D002465:Cell Movement / Q000235:genetics; D002522:Chlorocebus aethiops; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics* / Q000378:metabolism*; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D009097:Multienzyme Complexes; D016297:Mutagenesis, Site-Directed; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000378:metabolism*; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D011755:Pyrophosphatases; D018047:Receptors, Purinergic P1 / Q000502:physiology; D013329:Structure-Activity Relationship; D000099189:Lysophospholipase D
D005956:Glucose-6-Phosphate Isomerase; D006023:Glycoproteins; D009097:Multienzyme Complexes; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases; D018047:Receptors, Purinergic P1; D000099189:Lysophospholipase D