Water-Soluble Substrates of the Peptidoglycan-Modifying Enzyme O-Acetylpeptidoglycan Esterase (Ape1) from Neisseria gonorrheae

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• 作者：Timin Hadi ; John M. Pfeffer ; Anthony J. Clarke ; Martin E. Tanner
• 刊名：Journal of Organic Chemistry
• 出版年：2011
• 出版时间：February 18, 2011
• 年：2011
• 卷：76
• 期：4
• 页码：1118-1125
• 全文大小：852K
• 年卷期：v.76,no.4(February 18, 2011)
• ISSN：1520-6904

文摘

Peptidoglycan is the component of the bacterial cell wall that is essential for maintaining the shape and rigidity of the cell. As such, its polymeric structure, consisting of alternating units of N-acetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc), is also a target for the action of host defense enzymes, such as lysozymes. Many bacteria have developed methods of masking their cell wall from these environmental dangers through the addition of aglycon moieties that prevent recognition or sterically hinder the degradative action of exogenous enzymes that would otherwise prove detrimental to the cell. Peptidoglycan acetyl-transferases (Pat鈥檚) and O-acetylpeptidoglycan esterases (Ape鈥檚) are the enzymes responsible for the controlled addition and removal of acetate onto the C-6 hydroxyl group of MurNAc residues in peptidoglycan. Studies on Ape1, an O-acetylpeptidoglycan esterase found in Neisseria gonorrheae, have suggested that this enzyme is essential for bacterial viability and thus presents an attractive target for antibacterial design. Previous studies on Ape1 have been hindered by the fact that Ape1鈥檚 natural substrate is an insoluble polymer. In this paper we outline the design, synthesis, and testing of the water-soluble di- and monosaccharide substrate analogues 1 and 2. Both 1 and 2 serve as substrates of Ape1 with k_cat/K_M values of (5.1 卤 1.7) 脳 10³ M^鈭? s^鈭? and (3.1 卤 0.8) 脳 10³ M^鈭? s^鈭?, respectively. It was determined that the substitution of the GlcNAc residue in compound 1 with an O-benzyl group in compound 2 did not significantly decrease the enzyme鈥檚 affinity for the monosaccharide. These findings are important as they demonstrate that the catalytic prowess of Ape1 is not dependent on its binding to a polymeric substrate. This ensures that small molecule transition state/intermediate analogues can also capture the transition state binding energy of Ape1 and potentially serve as potent inhibitors. The synthetic route to compounds 1 and 2 could readily be modified to allow for the installation of a wide variety of functional groups at the MurNAc C-6 position in both the mono- and disaccharide scaffolds. This will serve as a general method for the construction of Ape1 substrates and inhibitors.