Microbial Biochemistry and Genetics Unit, OIIB
|
Molecular structure of sucrose-6P and its five phosphorylated isomers |
National Institute of Dental Reseach
BUILDING 30 ROOM 528
30 CONVENT DR, MSC 4352
BETHESDA MD 20892-4352
Phone: 301-496-4083
Fax: 301-402-0396
E-mail: jathomps@mail.nih.gov
Research Interests
This program examines the molecular basis for regulation of transport and dissimilation of carbohydrates and amino acids by oral microorganisms, and related species of lactic acid bacteria. Interests include the cloning and site - directed mutagenesis of genes, and the expression, purification and functional analysis of metabolic enzymes. Areas of specialization encompass the chemical and enzymatic syntheses of disaccharide phosphates, and significantly, the first preparation of the five phosphorylated isomers of sucrose (see photo). These novel derivatives are produced during membrane translocation via the bacterial phosphoenolpyruvate-dependent:sugar phosphotransferase system (PEP-PTS), and are hydrolyzed intracellularly by unique nucleotide and metal-dependent Family 4 glycosylhydrolases. In a multi-national collaboration, we have recently determined the crystal structure, active-site residues, and catalytic mechanism of the NAD+ and Mn2+-dependent phospho -alpha -glucosidase (GlvA) of Bacillus subtilis.
Education
B.Sc. Biochemistry (1st Class Honors), University of Leeds, England, 1960-1964
Ph.D. Biochemistry, University of Leeds, England, 1964-1967
Post-Doctoral Fellow, McGill University, Montreal, Canada, 1967-1971
Recent publications
Sackett, D.L., Ruvinov, S.B. and Thompson, J. 1999. N5 -(L-1-Carboxyethyl) ornithine synthase: physical and spectral characterization of the enzyme and its unusual low pKa fluorescent tyrosine residues. Protein Science 8: 2121-2129
Cisar, J. O., Xu, D-Q., Thompson, J., Swaim, W., Hu, L. and Kopecko, D. J. 2000. An alternative interpretation of nanobacteria- induced biomineralization. Proc. Natl. Acad. Sci. USA 97: 11511-11515
Yamamoto, H., Serizawa, M., Thompson, J. and Sekiguchi, J. 2001. Regulation of the glv operon in Bacillus subtilis: YfiA (GlvR) is a positive regulator of the operon, that is repressed through CcpA and cre. J. Bacteriol. 183: 5110- 5121
Thompson, J., Robrish, S. A., Pikis, A., Brust, A. and Lichtenthaler, F. W. 2001. Phosphorylation and metabolism of sucrose and its five linkage-isomeric alpha-D-glucosyl-D-fructoses by Klebsiella pneumoniae. 2001 Carbohydr. Res. 331:149-161
Thompson, J., Robrish, S. A., Immel, S., Lichtenthaler, F. W., Hall, B. G. and Pikis, A. 2001. Metabolism of sucrose and its five linkage-isomeric alpha-D-glucosyl-D-fructoses by Klebsiella pneumoniae: Participation and properties of sucrose-6-phosphate hydrolase and phospho-alpha-glucosidase. J. Biol. Chem. 276: 37415-37425
Thompson, J., Lichtenthaler, F.W., Peters, S. and Pikis, A. 2002. b -Glucoside kinase (BglK) from Klebsiella pneumoniae: Purification, properties, and preparative synthesis of 6- phospho-beta-glucosides. J. Biol. Chem. 277: 34310-34321
Thompson, J., Hess, S. and Pikis, A. 2004. Genes malh and pagl of Clostridium acetobutylicum ATCC 824 encode NAD+ and metal-dependent phospho-alpha-glucosidase(s). J. Biol. Chem. 279: 1553-1561
Yip, V.L.Y., Varrot, A., Davies, G.J., Rajan, S.S., Yang, X., Thompson, J., Anderson, W.F. and Withers, S.G. 2004. An unusual mechanism of glycoside hydrolysis involving redox and elimination steps by a Family 4 beta-glycosidase from Thermotoga maritima. J. Am. Chem. Soc. 126:8354-8355
Rajan, S.S., Yang, X., Collart, F., Yip, V.L.Y., Withers, S.G., Varrot, A., Thompson, J., Davies, G.J. and Anderson, W.F. 2004. Novel catalytic mechanism of glycoside hydrolysis based on the structure of an NAD+ /Mn2+ - dependent phospho-alpha-glucosidase from Bacillus subtilis. Structure 12:1619-1629.
Varrot, A., Yip, V.L.Y., Li, Y-S., Rajan, S.S., Yang, X., Anderson, W.F., Thompson, J., Withers, S.G., and Davies, G.J. 2005. NAD+ and metal-ion dependent hydrolysis by Family 4 glycosidases: structural insight into specificity for phospho-beta-glucosides. J. Mol. Biol. 346: 423-435.
Pikis, A., Hess, S., Arnold, I., Erni, B., and Thompson, J. 2006. Genetic requirements for growth of Escherichia coli K-12 on methyl-alpha-D-glucopyranoside and the five alpha-D-glucosyl-D-fructose isomers of sucrose. J. Biol. Chem. 281: 17900-17908.
Yip, V.L.Y., Thompson, J., and Withers, S.G. 2007. Mechanism of GlvA from Bacillus subtilis : A detailed kinetic analysis of a 6-phospho-alpha-glucosidase from glycoside hydrolase Family 4. Biochemistry 46: 9840-9852.
Thompson, J., Jakubovics, N., Abraham, B., Hess, S., and Pikis, A. 2008. The sim operon facilitates the transport and metabolism of sucrose isomers in Lactobacillus casei ATCC 334. J. Bacteriol. 190: 3362-3373