Development of metal-chelating inhibitors for the Class II fructose 1,6-bisphosphate (FBP) aldolase

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• 作者：Geneviè ; ve Labbé ; Anthony P. Krismanich ; Sarah de Groot ; Timothy Rasmusson ; Muhong Shang ; Matthew D.R. Brown ; Gary I. Dmitrienko ; J. Guy Guillemette ; ^{jguillem@uwaterloo.ca}
• 关键词：FBP aldolase ; Competitive slow-binding inhibitors
• 刊名：Journal of Inorganic Biochemistry
• 出版年：2012
• 期刊代码：53_01620134
• 类别：ch
• 出版时间：July, 2012
• 卷：112
• 期：Complete
• 页码：49-58
• 文件大小：1040 K

摘要

It has long been suggested that the essential and ubiquitous enzyme fructose 1,6-bisphosphate (FBP) aldolase could be a good drug target against bacteria and fungi, since lower organisms possess a metal-dependant (Class II) FBP aldolase, as opposed to higher organisms which possess a Schiff-base forming (Class I) FBP aldolase. We have tested the capacity of derivatives of the metal-chelating compound dipicolinic acid (DPA), as well a thiol-containing compound, to inhibit purified recombinant Class II FBP aldolases from Mycobacterium tuberculosis, Pseudomonas aeruginosa, Bacillus cereus, Bacillus anthracis, and from the Rice Blast causative agent Magnaporthe grisea. The aldolase from M. tuberculosis was the most sensitive to the metal-chelating inhibitors, with an IC₅₀ of 5.2 渭M with 2,3-dimercaptopropanesulfonate (DMPS) and 28 渭M with DPA. DMPS and the synthesized inhibitor 6-(phosphonomethyl)picolinic acid inhibited the enzyme in a time-dependent, competitive fashion, with second order rate constants of 273 and 270 M^鈭?#xA0;1 s^鈭?#xA0;1 respectively for the binding of these compounds to the M. tuberculosis aldolase's active site in the presence of the substrate FBP (K_M 27.9 渭M). The most potent first generation inhibitors were modeled into the active site of the M. tuberculosis aldolase structure, with results indicating that the metal chelators tested cannot bind the catalytic zinc in a bidentate fashion while it remains in its catalytic location, and that most enzyme-ligand interactions involve the phosphate binding pocket residues.