Glucosamine-6-phosphate synthase, a novel target for antifungal agents. Molecular modelling studies in drug design

• Autorzy: Wojciechowski M. , Milewski S. , Mazerski J. , Borowski E.
• Języki publikacji: EN

Abstrakty

EN

Fungal infections are a growing problem in contemporary medicine, yet only a few antifungal agents are used in clinical practice. In our laboratory we proposed the enzyme L-glutamine: D-fructose-6-phosphate amidotransferase (EC 2.6.1.16) as a new target for antifungals. The structure of this enzyme consists of two domains, N-terminal and C-terminal ones, catalysing glutamine hydrolysis and sugar-phosphate isomerisation, respectively. In our laboratory a series of potent selective inhibitors of GlcN-6-P synthase have been designed and synthesised. One group of these compounds, including the most studied N3-(4-methoxyfumaroyl)-l-2,3-diaminopropanoic acid (FMDP), behave like glutamine analogs acting as active-site-directed inactivators, blocking the N-terminal, glutamine-binding domain of the enzyme. The second group of GlcN-6-P synthase inhibitors mimic the transition state of the reaction taking place in the C-terminal sugar isomerising domain. Surprisingly, in spite of the fact that glutamine is the source of nitrogen for a number of enzymes it turned out that the glutamine analogue FMDP and its derivatives are selective against GlcN-6-P synthase and they do not block other enzymes, even belonging to the same family of glutamine amidotransferases. Our molecular modelling studies of this phenomenon revealed that even within the family of related enzymes substantial differences may exist in the geometry of the active site. In the case of the glutamine amidotransferase family the glutamine binding site of GlcN-6-P synthase fits a different region of the glutamine conformational space than other amidotransferases. Detailed analysis of the interaction pattern for the best known, so far, inhibitor of the sugar isomerising domain, namely 2-amino-2-deoxy-d-glucitol-6-phosphate (ADGP), allowed us to suggest changes in the structure of the inhibitor that should improve the interaction pattern. The novel ligand was designed and synthesised. Biological experiments confirmed our predictions. The new compound named ADMP is a much better inhibitor of glucosamine-6-phosphate synthase than ADGP.

Słowa kluczowe

EN

molecular modelling; drug design; glutamine aminotransferase; antifungal agent; glucosamine-6-phosphate synthase; fungal infection

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2005
• Tom: 52
• Numer: 3
• Opis fizyczny: p.647-653,fig.,ref.

Twórcy

autor

Wojciechowski M.

• Gdansk University of Technology, Gdansk, Poland

autor

Milewski S.

autor

Mazerski J.

autor

Borowski E.

Bibliografia

• Andruszkiewicz R, Chmara H, Milewski S, Borowski E (1986) Synthesis of N3-fumaramoyl-L-2,3-diaminopropanoic acid analogues, the irreversible inhibitors of glucosamine synthetase. Int J Pept Protein Res 27: 449–453.
• Andruszkiewicz R, Chmara H, Milewski S, Kasprzak L, Borowski E (1993) Structural determinants of inhibitory activity of N3-(4-methoxyfumaroyl)-L-2,3-diaminopropanoic acid towards glucosamine-6-phosphate synthase. Polish J Chem 67: 673–683.
• Andruszkiewicz R, Jedrzejczak R, Zieniawa T, Wojciechowski M, Borowski E (2000) N3-Oxoacyl derivatives of L-2,3-diaminopropanoic acid and their peptides; Novel inhibitors of glucosamine-6-phosphate synthase. J Enzyme Inhib 15: 429–441.
• Badet B, Vermoote P, Le Goffic F (1988) Glucosamine synthetase from Escherichia coli: kinetic mechanism and inhibition by N3-fumaroyl-L-2,3-diaminopropionic derivatives. Biochemistry 27: 2282–2287.
• Bates CJ, Adams WR, Handschumacher RE (1966) Control of the formation of uridine diphospho-N-acetyl-hexosamine and glycoprotein synthesis in rat liver. J Biol Chem 241: 1705–1712.
• Biosym Discover User Guide, version 2.9.5. (1995) Biosym/MSI, San Diego.
• Chmara H, Borowski E (1986) Bacteriolytic effect of cessation of glucosamine supply, induced by specific inhibition of glucosamine-6-phosphate synthetase. Acta Microbiol Pol 35: 15–27.
• Chmara H, Andruszkiewicz R, Borowski E (1984a) Inactivation of glucosamine-6-phosphate synthetase from Salmonella typhimurium LT2 SL 1027 by N-beta-fumarylcarboxyamido-L-2,3-diamino-propionic acid. Biochem Biophys Res Commun 120: 865–872.
• Chmara H, Zahner H, Borowski E, Milewski S (1984b) Inhibition of glucosamine-6-phosphate synthetase from bacteria by anticapsin. J Antibiot (Tokyo) 37: 652–658.
• Chmara H, Andruszkiewicz R, Borowski E (1985) Inactivation of glucosamine-6-phosphate synthetase from Salmonella typhimurium LT2 by fumaroyl diaminopropanoic acid derivatives, a novel group of glutamine analogs. Biochim Biophys Acta 870: 357–366.
• Crossley MJ, Stamford AW (1993) Concise, stereocontrolled synthesis of the C4 epimers of anticapsin and bacilysin: revision of the configurations of the natural products. Aust J Chem 46: 1443–1446.
• Isupov MN, Obmolova G, Butterworth S, Badet-Denisot MA, Badet B, Polikarpov I, Littlechild JA, Teplyakov A (1996) Substrate binding is required for assembly of the active conformation of the catalytic site in Ntn amidotransferases: Evidence from the 1.8 Å crystal structure of the glutaminase domain of glucosamine 6-phosphate synthase. Structure 4: 801–810.
• Kucharczyk N, Denisot MA, Le Goffic F, Badet B (1990)Glucosamine-6-phosphate synthase from Escherichia coli: determination of the mechanism of inactivation by N3-fumaroyl-L-2,3-diaminopropionic derivatives. Biochemistry 29: 3668–3676.
• Lindahl E, Hess B, van der Spoel D (2001) GROMACS 3.0: a package for molecular simulation and trajectory analysis. J Mol Model 7: 306–317.
• Madura JD, Briggs JM, Wade RC, Davis ME, Luty BA, Ilin A, Antosiewicz J, Gilson MK, Bagheri B, Scott LR, Mccammon JA (1995) Electrostatics and diffusion of molecules in solution: Simulations with the University of Houston Brownian Dynamics program. Comput Phys Commun 91: 57–95.
• Milewski S, Chmara H, Borowski E (1986) Antibiotic tetaine — a selective inhibitor of chitin and mannoprotein biosynthesis in Candida albicans. Arch Microbiol 145: 234–240.
• Milewski S, Chmara H, Andruszkiewicz R, Borowski E, Zaremba M, Borowski J (1988) Antifungal peptides with novel specific inhibitors of glucosamine 6-phosphate synthase. Drugs Exp Clin Res 14: 461–465.
• Milewski S, Chmara H, Andruszkiewicz R, Borowski E (1992) N3-haloacetyl derivatives of L-2,3-diaminopropanoic acid: novel inactivators of glucosamine-6-phosphate synthase. Biochim Biophys Acta 1115: 225–229.
• Morris GM, Goodsell DS, Halliday RS, Huey R, Hart WE, Belew EK, Olson AJ (1998) Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function. J Comput Chem 19: 1639–1662.
• Nicholls A, Sharp K, Honig B (1991) Protein folding and association: insight from the interfacial and thermodynamic properties of hydrocarbons. Protein Struct Funct Genet 11: 281–296.
• Smith RJ, Milewski S, Brown AJP, Gooday GW (1996) Isolation and characterization of the GFA1 gene encoding the glutamine:fructose-6-phosphate amidotransferase of Candida albicans. J Bacteriol 178: 2320–2327.
• Tarnowska M, Oldziej S, Liwo A, Grzonka Z, Borowski E (1992) Investigation of the inhibition pathway of glucosamine synthase by N3-(4-methoxyfumaroyl)-L-2,3-diaminopropanoic acid by semiempirical quantum mechanical and molecular mechanics methods. Eur Biophys J 21: 273–280.
• Tempczyk A, Tarnowska M, Liwo A (1989) A theoretical study of glucosamine synthase. Part I. Molecular mechanics calculations on substrate binding. Eur Biophys J 17: 201–210.
• Tempczyk A, Tarnowska M, Liwo A, Borowski E (1992) A theoretical study of glucosamine synthase. II. Combined quantum and molecular mechanics simulation of
• sullfydryl attack on the carboxyamide group. Eur Biophys J 21: 137–145.
• Teplyakov A (1998) Involvement of the C terminus in intramolecular nitrogen channeling in glucosamine 6-phosphate synthase: evidence from a 1.6 Å crystal structure of the isomerase domain. Structure 6: 1047–1055.
• Teplyakov A, Obmolova G, Badet-Denisot MA, Badet B (1999) The mechanism of sugar phosphate isomerization by glucosamine 6-phosphate synthase. Protein Sci 8: 596–602.
• Teplyakov A, Obmolova G, Badet-Denisot MA, Badet B (2001) Channeling of ammonia in glucosamine-6-phosphate synthase. J Mol Biol 313: 1093–1102.
• Teplyakov A, Leriche C, Obmolova G, Badet B, Badet- Denisot MA (2002) From Lobry de Bruyn to enzymecatalyzed ammonia channelling: molecular studies of D-glucosamine-6P synthase. Nat Prod Rep 19: 60–69.
• Wojciechowski M, Mazerski J, Borowski E (1995) Constrained search of conformational hyperspace of inactivators of glucosamine-6-phosphate synthase. J Enzym Inhib 10: 17–26.
• Wojciechowski M (1996) Molecular modelling of Glucosamine-6-phosphate synthase inhibition by glutamine analogs. PhD Thesis. Technical Universty of Gdansk, Gdansk.