HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 283, Issue 2, 792-801, January 11, 2008

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 283/2/792    most recent M705638200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Moriuchi, H. Articles by Watanabe, K. Search for Related Content PubMed PubMed Citation Articles by Moriuchi, H. Articles by Watanabe, K. Social Bookmarking                      What's this?

Molecular Characterization of a Novel Type of Prostamide/Prostaglandin F Synthase, Belonging to the Thioredoxin-like Superfamily^*

Hiroshi Moriuchi¹, Noriko Koda¹, Emiko Okuda-Ashitaka, Hiromi Daiyasu^¶, Kensuke Ogasawara, Hiroyuki Toh^||, Seiji Ito, David F. Woodward^**, and Kikuko Watanabe²

From the Division of Life Science, Graduate School of Integrated Science and Art, University of East Asia, 2-1 Ichinomiyagakuen, Shimonoseki, Yamaguchi 751-8503, Japan, Department of Medical Chemistry, Kansai Medical University, 10-15 Fumizono, Moriguchi, Osaka 570-8506, Japan, ^¶Center for Advanced Medical Engineering and Informatics, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan, ^||Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maedashi, Fukuoka 812-8582, Japan, and^**Department of Biological Sciences, Allergan, Inc., Irvine, California 92612

Prostaglandin F (PGF) ethanolamide (prostamide F) synthase, which catalyzed the reduction of prostamide H₂ to prostamide F₂, was found in mouse and swine brain. The enzyme was purified from swine brain, and its amino acid sequence was defined. The mouse enzyme consisted of a 603-bp open reading frame coding for a 201-amino acid polypeptide with a molecular weight of 21,669. The amino acid sequence placed the enzyme in the thioredoxin-like superfamily with Cys⁴⁴ being the active site. The enzyme expressed in Escherichia coli as well as the native enzyme catalyzed not only the reduction of prostamide H₂ to prostamide F₂ but also that of PGH₂ to PGF₂. The V_max and K_m values for prostamide H₂ were about 0.25 µmol/min·mg of protein and 7.6 µM, respectively, and those for PGH₂ were about 0.69 µmol/min·mg of protein and 6.9 µM, respectively. Neither PGE₂ nor PGD₂ served as a substrate for this synthase. Based on these data, we named the enzyme prostamide/PGF synthase. Although the enzyme showed a broad specificity for reductants, reduced thioredoxin preferentially served as a reducing equivalent donor for this enzyme. Moreover, Northern and Western blot analyses in addition to the prostamide F synthase activity showed that the enzyme was mainly distributed in the brain and spinal cord, and the immunohistochemical study in the spinal cord showed that the enzyme was found mainly in the cytosol. These results suggest that prostamide/PGF synthase may play an important functional role in the central nervous system.

Received for publication, July 10, 2007 , and in revised form, November 8, 2007.

^* 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 experimental procedures, Tables I and II, and Figs. 1–4.

The nucleotide sequence(s) reported in this paper has been submitted to the DDBJ/GenBankTM/EBI Data Bank with accession number(s) AB329665.

¹ These authors contributed equally to this work.

² To whom correspondence should be addressed. Tel.: 81-832-57-5152; Fax: 81-832-57-5152; E-mail: watanabe{at}toua-u.ac.jp.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. W. Buczynski, D. S. Dumlao, and E. A. Dennis Thematic Review Series: Proteomics. An integrated omics analysis of eicosanoid biology J. Lipid Res., June 1, 2009; 50(6): 1015 - 1038. [Abstract] [Full Text] [PDF]