Cyclic phosphatidic acid is produced by autotaxin in blood.
Cyclic phosphatidic acid (cPA), an analog of lysophosphatidic acid (LPA), was previously identified in human serum. Although cPA possesses distinct physiological activities not elicited by LPA, its biochemical origins have scarcely been studied. In the present study, we assayed cPA formation from lysophosphatidylcholine in fetal bovine serum and found significant activity of transphosphatidylation that generated cPA. The cPA-producing enzyme was purified from fetal bovine serum using five chromatographic steps yielding a 100-kDa protein with cPA biosynthetic activity. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of its tryptic peptides revealed that the enzyme shared identical fragments with human autotaxin, a serum lysophospholipase D that produces LPA. Western blot analysis demonstrated that the 100-kDa protein was specifically recognized by an anti-human autotaxin antibody. Moreover, recombinant rat autotaxin was found to generate cPA in addition to LPA. No significant cPA- or LPA-producing activity was detected in autotaxin-depleted serum from bovine or human prepared by immunoprecipitation with an anti-autotaxin monoclonal antibody. These results indicate that the generation of cPA and LPA in serum is mainly attributed to autotaxin.
Tsuda
Satomi
S
Department of Biology, Faculty of Science, Ochanomizu University, Tokyo 112-8610, Japan.
Okudaira
Shinichi
S
Moriya-Ito
Keiko
K
Shimamoto
Chie
C
Tanaka
Masayuki
M
Aoki
Junken
J
Arai
Hiroyuki
H
Murakami-Murofushi
Kimiko
K
Kobayashi
Tetsuyuki
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001769:Blood / Q000378:metabolism*; D002417:Cattle; D004986:Ether / Q000378:metabolism; D006801:Humans; D008244:Lysophosphatidylcholines / Q000378:metabolism; D008246:Lysophospholipids / Q000096:biosynthesis / Q000737:chemistry; D008670:Metals / Q000378:metabolism; D009097:Multienzyme Complexes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D043264:Phosphodiesterase I / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011755:Pyrophosphatases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012965:Sodium Chloride / Q000378:metabolism; D000099189:Lysophospholipase D
D000911:Antibodies, Monoclonal; D004986:Ether; D008244:Lysophosphatidylcholines; D008246:Lysophospholipids; D008670:Metals; D009097:Multienzyme Complexes; D010446:Peptide Fragments; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases; D011994:Recombinant Proteins; D012965:Sodium Chloride; D000099189:Lysophospholipase D; C032881:lysophosphatidic acid