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J. Biol. Chem., Vol. 283, Issue 12, 7776-7789, March 21, 2008

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Murine and Human Autotaxin , β, and Isoforms

GENE ORGANIZATION, TISSUE DISTRIBUTION, AND BIOCHEMICAL CHARACTERIZATION^*

Adeline Giganti, Marianne Rodriguez, Benjamin Fould, Natacha Moulharat, Francis Cogé, Pascale Chomarat, Jean-Pierre Galizzi, Philippe Valet^¶, Jean-Sébastien Saulnier-Blache^¶, Jean A. Boutin¹, and Gilles Ferry

From the Pharmacologie Moléculaire et Cellulaire and Pharmacologie et Physiopathologie Moléculaire, Institut de Recherches Servier, 78290 Croissy-sur-Seine and ^¶INSERM U858/I2MR, Department of Metabolism and Obesity, Team 3, 1 Avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France

Autotaxin is a type II ectonucleotide pyrophosphate phosphodiesterase enzyme. It has been recently discovered that it also has a lysophospholipase D activity. This enzyme probably provides most of the extracellular lysophosphatidic acid from lysophosphatidylcholine. The cloning and tissue distribution of the three isoforms (imaginatively called , β, and ) from human and mouse are reported in this study, as well as their tissue distribution by PCR in the human and mouse. The fate of the isoform from human was also studied after purification and using mass spectrometry. Indeed, this particular isoform expresses the intron 12 in which a cleavage site is present, leading to a rapid catabolism of the isoform. For the human isoform and the total autotaxin mRNA expression, quantitative PCR is presented in 21 tissues. The isoforms were expressed in two different hosts, insect cells and Chinese hamster ovary cells, and were highly purified. The characteristics of the six purified isoforms (pH and temperature dependence, K_m and V_max values, and their dependence on metal ions) are presented in this study. Their sensitivity to a small molecule inhibitor, hypericin, is also shown. Finally, the specificity of the isoforms toward a large family of lysophosphatidylcholines is reported. This study is the first complete description of the reported autotaxin isoforms.

Received for publication, October 22, 2007 , and in revised form, December 31, 2007.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) EU131009 [GenBank] , EU131010 [GenBank] , and EU131011.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. 1 and Tables S1–S3.

¹ To whom correspondence should be addressed: Pharmacologie Moléculaire et Cellulaire, Institut de Recherches SERVIER, 125, Chemin de Ronde, 78290 Croissy-sur-Seine, France. Tel.: 33-1-55-72-27-48; Fax: 33-1-55-72-28-10; E-mail: jean.boutin{at}fr.netgrs.com.

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