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The catalytic mechanism of galactose mutarotase.

Thoden JB, Kim J, Raushel FM, Holden HM
Protein Sci 12 1051-9 (2003)
Related entries: 1ns2, 1ns4, 1ns7, 1ns8, 1nsm, 1nsr, 1nss, 1nsu, 1nsv, 1nsx, 1nsz

Cited: 22 times
EuropePMC logo PMID: 12717027

Abstract

Galactose mutarotase catalyzes the first step in normal galactose metabolism by catalyzing the conversion of beta-D-galactose to alpha-D-galactose. The structure of the enzyme from Lactococcus lactis was recently solved in this laboratory and shown to be topologically similar to domain 5 of beta-galactosidase. From this initial X-ray analysis, four amino acid residues were demonstrated to be intimately involved in sugar binding to the protein: His 96, His 170, Asp 243, and Glu 304. Here we present a combined X-ray crystallographic and kinetic analysis designed to examine the role of these residues in the reaction mechanism of the enzyme. For this investigation, the following site-directed mutant proteins were prepared: H96N, H170N, D243N, D243A, E304Q, and E304A. All of the structures of these proteins, complexed with either glucose or galactose, were solved to a nominal resolution of 1.95 A or better, and their kinetic parameters were measured against D-galactose, D-glucose, L-arabinose, or D-xylose. From these studies, it can be concluded that Glu 304 and His 170 are critical for catalysis and that His 96 and Asp 243 are important for proper substrate positioning within the active site. Specifically, Glu 304 serves as the active site base to initiate the reaction by removing the proton from the C-1 hydroxyl group of the sugar substrate and His 170 functions as the active site acid to protonate the C-5 ring oxygen.

Articles - 1ns0 mentioned but not cited (3)

  1. The catalytic mechanism of galactose mutarotase. Thoden JB, Kim J, Raushel FM, Holden HM. Protein Sci 12 1051-1059 (2003)
  2. Automated identification of protein-ligand interaction features using Inductive Logic Programming: a hexose binding case study. A Santos JC, Nassif H, Page D, Muggleton SH, E Sternberg MJ. BMC Bioinformatics 13 162 (2012)
  3. An Inductive Logic Programming Approach to Validate Hexose Binding Biochemical Knowledge. Nassif H, Al-Ali H, Khuri S, Keirouz W, Page D. Inductive Log Program 5989 149-165 (2010)


Reviews citing this publication (2)

  1. Structure and function of enzymes of the Leloir pathway for galactose metabolism. Holden HM, Rayment I, Thoden JB. J Biol Chem 278 43885-43888 (2003)
  2. Therapeutic Monosaccharides: Looking Back, Moving Forward. Sosicka P, Ng BG, Freeze HH. Biochemistry 59 3064-3077 (2020)

Articles citing this publication (17)

  1. The generation of new protein functions by the combination of domains. Bashton M, Chothia C. Structure 15 85-99 (2007)
  2. A rapid coarse residue-based computational method for x-ray solution scattering characterization of protein folds and multiple conformational states of large protein complexes. Yang S, Park S, Makowski L, Roux B. Biophys J 96 4449-4463 (2009)
  3. KdgF, the missing link in the microbial metabolism of uronate sugars from pectin and alginate. Hobbs JK, Lee SM, Robb M, Hof F, Barr C, Abe KT, Hehemann JH, McLean R, Abbott DW, Boraston AB. Proc Natl Acad Sci U S A 113 6188-6193 (2016)
  4. UDPgalactose 4-epimerase from Saccharomyces cerevisiae. A bifunctional enzyme with aldose 1-epimerase activity. Majumdar S, Ghatak J, Mukherji S, Bhattacharjee H, Bhaduri A. Eur J Biochem 271 753-759 (2004)
  5. The central cavity from the (alpha/alpha)6 barrel structure of Anabaena sp. CH1 N-acetyl-D-glucosamine 2-epimerase contains two key histidine residues for reversible conversion. Lee YC, Wu HM, Chang YN, Wang WC, Hsu WH. J Mol Biol 367 895-908 (2007)
  6. Crystal structure of YihS in complex with D-mannose: structural annotation of Escherichia coli and Salmonella enterica yihS-encoded proteins to an aldose-ketose isomerase. Itoh T, Mikami B, Hashimoto W, Murata K. J Mol Biol 377 1443-1459 (2008)
  7. Structure-based functional annotation: yeast ymr099c codes for a D-hexose-6-phosphate mutarotase. Graille M, Baltaze JP, Leulliot N, Liger D, Quevillon-Cheruel S, van Tilbeurgh H. J Biol Chem 281 30175-30185 (2006)
  8. Structural insights into the epimerization of β-1,4-linked oligosaccharides catalyzed by cellobiose 2-epimerase, the sole enzyme epimerizing non-anomeric hydroxyl groups of unmodified sugars. Fujiwara T, Saburi W, Matsui H, Mori H, Yao M. J Biol Chem 289 3405-3415 (2014)
  9. RhaU of Rhizobium leguminosarum is a rhamnose mutarotase. Richardson JS, Carpena X, Switala J, Perez-Luque R, Donald LJ, Loewen PC, Oresnik IJ. J Bacteriol 190 2903-2910 (2008)
  10. Induction of the galactose enzymes in Escherichia coli is independent of the C-1-hydroxyl optical configuration of the inducer D-galactose. Lee SJ, Lewis DE, Adhya S. J Bacteriol 190 7932-7938 (2008)
  11. Potential roles of inorganic phosphate on the progression of initially bound glucopyranose toward the nonenzymatic glycation of human hemoglobin: mechanistic diversity and impacts on site selectivity. Smith BA, Mottishaw CR, Hendricks AJ, Mitchell J, Becker S, Ropski PS, Park B, Finkbeiner-Caufield M, Garay-Nontol B, Holman RW, Rodnick KJ. Cogent Biol 4 1425196 (2018)
  12. Crystal structures and enzyme mechanisms of a dual fucose mutarotase/ribose pyranase. Lee KH, Ryu KS, Kim MS, Suh HY, Ku B, Song YL, Ko S, Lee W, Oh BH. J Mol Biol 391 178-191 (2009)
  13. Deciphering the function of an ORF: Salmonella enterica DeoM protein is a new mutarotase specific for deoxyribose. Assairi L, Bertrand T, Ferdinand J, Slavova-Azmanova N, Christensen M, Briozzo P, Schaeffer F, Craescu CT, Neuhard J, Bârzu O, Gilles AM. Protein Sci 13 1295-1303 (2004)
  14. Crystallization and preliminary X-ray diffraction analysis of the putative aldose 1-epimerase YeaD from Escherichia coli. You W, Qiu X, Zhang Y, Ma J, Gao Y, Zhang X, Niu L, Teng M. Acta Crystallogr Sect F Struct Biol Cryst Commun 66 951-953 (2010)
  15. Characterisation and expression of a gene encoding a mutarotase from the fungus Rhizopus nigricans. Vilfan T, Cresnar B, Fournier D, Stojan J, Breskvar K. FEMS Microbiol Lett 235 101-108 (2004)
  16. Comparative Genomic Analyses of Virulence and Antimicrobial Resistance in Citrobacter werkmanii, an Emerging Opportunistic Pathogen. Aguirre-Sánchez JR, Quiñones B, Ortiz-Muñoz JA, Prieto-Alvarado R, Vega-López IF, Martínez-Urtaza J, Lee BG, Chaidez C. Microorganisms 11 2114 (2023)
  17. The hypergonadotropic hypogonadism conundrum of classic galactosemia. Derks B, Rivera-Cruz G, Hagen-Lillevik S, Vos EN, Demirbas D, Lai K, Treacy EP, Levy HL, Wilkins-Haug LE, Rubio-Gozalbo ME, Berry GT. Hum Reprod Update 29 246-258 (2023)


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