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

J. Biol. Chem., Vol. 279, Issue 52, 54687-54694, December 24, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/52/54687    most recent M411023200v1 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 Proudfoot, M. Articles by Yakunin, A. F. Search for Related Content PubMed PubMed Citation Articles by Proudfoot, M. Articles by Yakunin, A. F. Social Bookmarking                      What's this?

General Enzymatic Screens Identify Three New Nucleotidases in Escherichia coli

BIOCHEMICAL CHARACTERIZATION OF SurE, YfbR, AND YjjG^*

Michael Proudfoot, Ekaterina Kuznetsova, Greg Brown, Narayana N. Rao¶, Masanari Kitagawa||**, Hirotada Mori**, Alexei Savchenko, and Alexander F. Yakunin

From the Banting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, Canada, the ^¶School of Medicine, Stanford University, Stanford, California 94305-5307, the ^||Dragon Genomics Center, Takara Bio Incorporated, 7870-15, Sakura-cho, Yokkaichi, Mie 512-1211, Japan, and the Research and Education Center for Genetic Information, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan

To find proteins with nucleotidase activity in Escherichia coli, purified unknown proteins were screened for the presence of phosphatase activity using the general phosphatase substrate p-nitrophenyl phosphate. Proteins exhibiting catalytic activity were then assayed for nucleotidase activity against various nucleotides. These screens identified the presence of nucleotidase activity in three uncharacterized E. coli proteins, SurE, YfbR, and YjjG, that belong to different enzyme superfamilies: SurE-like family, HD domain family (YfbR), and haloacid dehalogenase (HAD)-like superfamily (YjjG). The phosphatase activity of these proteins had a neutral pH optimum (pH 7.0–8.0) and was strictly dependent on the presence of divalent metal cations (SurE: Mn²⁺ > Co²⁺ > Ni²⁺ > Mg²⁺; YfbR: Co²⁺ > Mn²⁺ > Cu²⁺; YjjG: Mg²⁺ > Mn²⁺ > Co²⁺). Further biochemical characterization of SurE revealed that it has a broad substrate specificity and can dephosphorylate various ribo- and deoxyribonucleoside 5'-monophosphates and ribonucleoside 3'-monophosphates with highest affinity to 3'-AMP. SurE also hydrolyzed polyphosphate (exopolyphosphatase activity) with the preference for short-chain-length substrates (P_20–25). YfbR was strictly specific to deoxyribonucleoside 5'-monophosphates, whereas YjjG showed narrow specificity to 5'-dTMP, 5'-dUMP, and 5'-UMP. The three enzymes also exhibited different sensitivities to inhibition by various nucleoside di- and triphosphates: YfbR was equally sensitive to both di- and triphosphates, SurE was inhibited only by triphosphates, and YjjG was insensitive to these effectors. The differences in their sensitivities to nucleotides and their varied substrate specificities suggest that these enzymes play unique functions in the intracellular nucleotide metabolism in E. coli.

Received for publication, September 24, 2004 , and in revised form, October 7, 2004.

^* This work was supported in part by Genome Canada, the Ontario Research and Development Challenge Fund, the Protein Structure Initiative of the National Institutes of Health (Midwest Center for Structural Genomics) Grant GM62414). 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.

Both authors contributed equally to this work.

^** Supported by CREST (Japan Science and Technology) and the Inamori Foundation.

To whom correspondence should be addressed. Tel.: 416-946-0075; Fax: 416-978-8528; E-mail: a.iakounine{at}utoronto.ca.

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

This article has been cited by other articles:

				H. S. Lee, Y. J. Kim, J.-H. Lee, and S. G. Kang Identification and Characterization of Inorganic Pyrophosphatase and PAP Phosphatase from Thermococcus onnurineus NA1 J. Bacteriol., May 15, 2009; 191(10): 3415 - 3419. [Abstract] [Full Text] [PDF]

				S. N. Lindner, S. Knebel, H. Wesseling, S. M. Schoberth, and V. F. Wendisch Exopolyphosphatases PPX1 and PPX2 from Corynebacterium glutamicum Appl. Envir. Microbiol., May 15, 2009; 75(10): 3161 - 3170. [Abstract] [Full Text] [PDF]

				G. Brown, A. Singer, V. V. Lunin, M. Proudfoot, T. Skarina, R. Flick, S. Kochinyan, R. Sanishvili, A. Joachimiak, A. M. Edwards, et al. Structural and Biochemical Characterization of the Type II Fructose-1,6-bisphosphatase GlpX from Escherichia coli J. Biol. Chem., February 6, 2009; 284(6): 3784 - 3792. [Abstract] [Full Text] [PDF]

				M. M. Adams, M. R. Gomez-Garcia, A. R. Grossman, and D. Bhaya Phosphorus Deprivation Responses and Phosphonate Utilization in a Thermophilic Synechococcus sp. from Microbial Mats J. Bacteriol., December 15, 2008; 190(24): 8171 - 8184. [Abstract] [Full Text] [PDF]

				H. S. Lee, Y. Cho, J.-H. Lee, and S. G. Kang Novel Monofunctional Histidinol-Phosphate Phosphatase of the DDDD Superfamily of Phosphohydrolases J. Bacteriol., April 1, 2008; 190(7): 2629 - 2632. [Abstract] [Full Text] [PDF]

				B. Weiss The Deoxycytidine Pathway for Thymidylate Synthesis in Escherichia coli J. Bacteriol., November 1, 2007; 189(21): 7922 - 7926. [Abstract] [Full Text] [PDF]

				L. Saskova, L. Novakova, M. Basler, and P. Branny Eukaryotic-Type Serine/Threonine Protein Kinase StkP Is a Global Regulator of Gene Expression in Streptococcus pneumoniae J. Bacteriol., June 1, 2007; 189(11): 4168 - 4179. [Abstract] [Full Text] [PDF]

				B. Weiss YjjG, a dUMP Phosphatase, Is Critical for Thymine Utilization by Escherichia coli K-12 J. Bacteriol., March 1, 2007; 189(5): 2186 - 2189. [Abstract] [Full Text] [PDF]

				E. Kuznetsova, M. Proudfoot, C. F. Gonzalez, G. Brown, M. V. Omelchenko, I. Borozan, L. Carmel, Y. I. Wolf, H. Mori, A. V. Savchenko, et al. Genome-wide Analysis of Substrate Specificities of the Escherichia coli Haloacid Dehalogenase-like Phosphatase Family J. Biol. Chem., November 24, 2006; 281(47): 36149 - 36161. [Abstract] [Full Text] [PDF]

				M. Kitagawa, T. Ara, M. Arifuzzaman, T. Ioka-Nakamichi, E. Inamoto, H. Toyonaga, and H. Mori Complete set of ORF clones of Escherichia coli ASKA library (A Complete Set of E. coli K-12 ORF Archive): Unique Resources for Biological Research DNA Res, January 1, 2005; 12(5): 291 - 299. [Abstract] [Full Text] [PDF]