John Thompson, Ph.D.

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 (1^st 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.