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

This Article Full Text (PDF) 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Giegel, D. A. Articles by Massey, V. Search for Related Content PubMed PubMed Citation Articles by Giegel, D. A. Articles by Massey, V. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 265, Issue 12, 6626-6632, Apr, 1990

L-lactate 2-monooxygenase from Mycobacterium smegmatis. Cloning, nucleotide sequence, and primary structure homology within an enzyme family

DA Giegel, CH Williams Jr and V Massey
Department of Biological Chemistry, University of Michigan, Ann Arbor.

L-Lactate 2-monooxygenase catalyzes the oxidation of L-lactate to acetate and carbon dioxide. The catalytic mechanism has been extensively investigated but very little is known about which amino acid residues may play a role in catalysis. As a first step toward this goal, the gene for this protein from Mycobacterium smegmatis has been cloned and sequenced. Peptide sequencing data for L-lactate 2- monooxygenase was used to construct three sets of fully redundant tetradecamer oligonucleotide probes, which were hybridized to restriction-digested M. smegmatis DNA. An approximately 3-kilobase pair PstI fragment hybridized with two of the probes. This region was subsequently isolated and cloned into Escherichia coli. From this size- fractionated gene bank, a 3.1-kilobase pair genomic DNA fragment was isolated by colony hybridization to two of the oligonucleotide probes. The complete gene for L-lactate 2-monooxygenase was contained on this fragment as shown by DNA sequencing of the whole insert. The DNA sequence codes for a mature protein that is 393 amino acids in length with a subunit molecular weight of 43,072 (including the FMN). The protein sequence shows impressive homology with the primary structures of two mechanistically related proteins, yeast flavocytochrome b2 (Lederer, F., Cortial, S., Becam, A.-M., Haumont, P.-Y., and Perez, L. (1985) Eur. J. Biochem. 152, 419-428; Guiard, B. (1985) EMBO J. 4, 3265- 3272) and spinach glycolate oxidase (Volkita, M., and Somerville, C. R. (1987) J. Biol. Chem. 262, 15825-15828; Cederlund, E., Lindqvist, Y., Soderlund, G., Branden, C.-I., and Jornvall, H. (1988) Eur. J. Biochem. 173, 523-530). For each residue proposed from the crystal structure of glycolate oxidase to be involved in catalysis (Lindqvist, Y., and Branden, C.-I. (1989) J. Biol. Chem. 264, 3624-3628), an identical residue was found in a homologous position in lactate oxidase. Furthermore, most of these residues occur in regions whose sequences are highly conserved between lactate oxidase, flavocytochrome b2, and glycolate oxidase.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				N. L. Ferreira, D. Labbe, F. Monot, F. Fayolle-Guichard, and C. W. Greer Genes involved in the methyl tert-butyl ether (MTBE) metabolic pathway of Mycobacterium austroafricanum IFP 2012. Microbiology, May 1, 2006; 152(Pt 5): 1361 - 1374. [Abstract] [Full Text] [PDF]

				K. Francis, B. Russell, and G. Gadda Involvement of a Flavosemiquinone in the Enzymatic Oxidation of Nitroalkanes Catalyzed by 2-Nitropropane Dioxygenase J. Biol. Chem., February 18, 2005; 280(7): 5195 - 5204. [Abstract] [Full Text] [PDF]

				B. G. Kelly, D. M. Wall, C. A. Boland, and W. G. Meijer Isocitrate lyase of the facultative intracellular pathogen Rhodococcus equi Microbiology, March 1, 2002; 148(3): 793 - 798. [Abstract] [Full Text] [PDF]

				A. Gibello, M. D. Collins, L. Domínguez, J. F. Fernández-Garayzábal, and P. T. Richardson Cloning and Analysis of the L-Lactate Utilization Genes from Streptococcus iniae Appl. Envir. Microbiol., October 1, 1999; 65(10): 4346 - 4350. [Abstract] [Full Text]

				S. A. Sanders, C. H. Williams Jr., and V. Massey The Roles of Two Amino Acid Residues in the Active Site of L-Lactate Monooxygenase. MUTATION OF ARGININE 187 TO METHIONINE AND HISTIDINE 240 TO GLUTAMINE J. Biol. Chem., August 6, 1999; 274(32): 22289 - 22295. [Abstract] [Full Text] [PDF]

				W. Sun, C. H. Williams Jr., and V. Massey The Role of Glycine 99 in L-Lactate Monooxygenase from Mycobacterium smegmatis J. Biol. Chem., October 24, 1997; 272(43): 27065 - 27076. [Abstract] [Full Text] [PDF]

				K. Yorita, K. Janko, K. Aki, S. Ghisla, B. A. Palfey, and V. Massey On the reaction mechanism of L-lactate oxidase: Quantitative structure-activity analysis of the reaction with para-substituted L-mandelates PNAS, September 2, 1997; 94(18): 9590 - 9595. [Abstract] [Full Text] [PDF]

				W. Sun, C. H. Williams Jr., and V. Massey Site-directed Mutagenesis of Glycine 99to Alanine in L-Lactate Monooxygenase from Mycobacterium smegmatis J. Biol. Chem., July 19, 1996; 271(29): 17226 - 17233. [Abstract] [Full Text] [PDF]