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

J. Biol. Chem., Vol. 279, Issue 44, 45728-45736, October 29, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/44/45728    most recent M408716200v1 Alert me when this article is cited 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 Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kotake, T. Articles by Tsumuraya, Y. Search for Related Content PubMed PubMed Citation Articles by Kotake, T. Articles by Tsumuraya, Y. Social Bookmarking                      What's this?

UDP-sugar Pyrophosphorylase with Broad Substrate Specificity Toward Various Monosaccharide 1-Phosphates from Pea Sprouts^*

Toshihisa Kotake, Daisuke Yamaguchi, Hiroshi Ohzono, Sachiko Hojo, Satoshi Kaneko¶, Hide-ki Ishida||, and Yoichi Tsumuraya

From the Department of Biochemistry and Molecular Biology, Faculty of Science, Saitama University, 255 Shimo-okubo, Sakura-ku, Saitama 338-8570, Japan, ^¶Biological Function Division, National Food Research Institute, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8642, Japan, and the ^||Noguchi Institute, 1-8-1 Kaga, Itabashi-ku, Tokyo 173-0003, Japan

UDP-sugars, activated forms of monosaccharides, are synthesized through de novo and salvage pathways and serve as substrates for the synthesis of polysaccharides, glycolipids, and glycoproteins in higher plants. A UDP-sugar pyrophosphorylase, designated PsUSP, was purified about 1,200-fold from pea (Pisum sativum L.) sprouts by conventional chromatography. The apparent molecular mass of the purified PsUSP was 67,000 Da. The enzyme catalyzed the formation of UDP-Glc, UDP-Gal, UDP-glucuronic acid, UDP-L-arabinose, and UDP-xylose from respective monosaccharide 1-phosphates in the presence of UTP as a co-substrate, indicating that the enzyme has broad substrate specificity toward monosaccharide 1-phosphates. Maximum activity of the enzyme occurred at pH 6.5–7.5, and at 45 °C in the presence of 2 mM Mg²⁺. The apparent K_m values for Glc 1-phosphate and L-arabinose 1-phosphate were 0.34 and 0.96 mM, respectively. PsUSP cDNA was cloned by reverse transcriptase-PCR. PsUSP appears to encode a protein with a molecular mass of 66,040 Da (600 amino acids) and possesses a uridine-binding site, which has also been found in a human UDP-N-acetylhexosamine pyrophosphorylase. Phylogenetic analysis revealed that PsUSP can be categorized in a group together with homologues from Arabidopsis and rice, which is distinct from the UDP-Glc and UDP-N-acetylhexosamine pyrophosphorylase groups. Recombinant PsUSP expressed in Escherichia coli catalyzed the formation of UDP-sugars from monosaccharide 1-phosphates and UTP with efficiency similar to that of the native enzyme. These results indicate that the enzyme is a novel type of UDP-sugar pyrophosphorylase, which catalyzes the formation of various UDP-sugars at the end of salvage pathways in higher plants.

Received for publication, July 30, 2004 , and in revised form, August 23, 2004.

^* 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 nucleotide sequence(s) reported in this paper has been submitted to the DDBJ/GenBank^TM/EBI Data Bank with accession number(s) AB178642.

To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, Faculty of Science, Saitama University, 255 Shimo-okubo, Sakura-ku, Saitama 338-8570, Japan. Tel.: 81-48-858-3955; Fax: 81-48-858-3384; E-mail: kotake{at}molbiol.saitama-u.ac.jp.

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

This article has been cited by other articles:

				T. Kotake, S. Hojo, N. Tajima, K. Matsuoka, T. Koyama, and Y. Tsumuraya A Bifunctional Enzyme with L-Fucokinase and GDP-L-fucose Pyrophosphorylase Activities Salvages Free L-Fucose in Arabidopsis J. Biol. Chem., March 28, 2008; 283(13): 8125 - 8135. [Abstract] [Full Text] [PDF]

				H. D. Coleman, T. Canam, K.-Y. Kang, D. D. Ellis, and S. D. Mansfield Over-expression of UDP-glucose pyrophosphorylase in hybrid poplar affects carbon allocation J. Exp. Bot., December 1, 2007; 58(15-16): 4257 - 4268. [Abstract] [Full Text] [PDF]

				R. Moretti and J. S. Thorson Enhancing the Latent Nucleotide Triphosphate Flexibility of the Glucose-1-phosphate Thymidylyltransferase RmlA J. Biol. Chem., June 8, 2007; 282(23): 16942 - 16947. [Abstract] [Full Text] [PDF]

				T. Oka, T. Nemoto, and Y. Jigami Functional Analysis of Arabidopsis thaliana RHM2/MUM4, a Multidomain Protein Involved in UDP-D-glucose to UDP-L-rhamnose Conversion J. Biol. Chem., February 23, 2007; 282(8): 5389 - 5403. [Abstract] [Full Text] [PDF]

				N. Dai, M. Petreikov, V. Portnoy, N. Katzir, D. M. Pharr, and A. A. Schaffer Cloning and Expression Analysis of a UDP-Galactose/Glucose Pyrophosphorylase from Melon Fruit Provides Evidence for the Major Metabolic Pathway of Galactose Metabolism in Raffinose Oligosaccharide Metabolizing Plants Plant Physiology, September 1, 2006; 142(1): 294 - 304. [Abstract] [Full Text] [PDF]

				T. Kotake, K. Tsuchiya, T. Aohara, T. Konishi, S. Kaneko, K. Igarashi, M. Samejima, and Y. Tsumuraya An {alpha}-L-arabinofuranosidase/{beta}-D-xylosidase from immature seeds of radish (Raphanus sativus L.) J. Exp. Bot., July 1, 2006; 57(10): 2353 - 2362. [Abstract] [Full Text] [PDF]

				A. Ashikov, F. Routier, J. Fuhlrott, Y. Helmus, M. Wild, R. Gerardy-Schahn, and H. Bakker The Human Solute Carrier Gene SLC35B4 Encodes a Bifunctional Nucleotide Sugar Transporter with Specificity for UDP-Xylose and UDP-N-Acetylglucosamine J. Biol. Chem., July 22, 2005; 280(29): 27230 - 27235. [Abstract] [Full Text] [PDF]

				T. Kotake, S. Dina, T. Konishi, S. Kaneko, K. Igarashi, M. Samejima, Y. Watanabe, K. Kimura, and Y. Tsumuraya Molecular Cloning of a {beta}-Galactosidase from Radish That Specifically Hydrolyzes {beta}-(1->3)- and {beta}-(1->6)-Galactosyl Residues of Arabinogalactan Protein Plant Physiology, July 1, 2005; 138(3): 1563 - 1576. [Abstract] [Full Text] [PDF]

				C.-W. Chang, J. L. Moseley, D. Wykoff, and A. R. Grossman The LPB1 Gene Is Important for Acclimation of Chlamydomonas reinhardtii to Phosphorus and Sulfur Deprivation Plant Physiology, May 1, 2005; 138(1): 319 - 329. [Abstract] [Full Text] [PDF]