Sanford-Burnham Medical Research Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, United States.
Int J Biochem Cell Biol. 2010 Jun;42(6):987-95. doi: 10.1016/j.biocel.2010.02.013. Epub 2010 Mar 1.
Furin and related proprotein convertases cleave the multibasic motifs R-X-R/K/X-R in the precursor proteins and, as a result, transform the latent proproteins into biologically active proteins and peptides. Furin is present both in the intracellular secretory pathway and at the cell surface. Intracellular furin processes its multiple normal cellular targets in the Golgi and secretory vesicle compartments while cell-surface furin appears to be essential only for the processing of certain pathogenic proteins and, importantly, anthrax. To design potent, safe and selective inhibitors of furin, we evaluated the potency and selectivity of the derivatized peptidic inhibitors modeled from the extended furin cleavage sequence of avian influenza A H5N1. We determined that the N- and C-terminal modifications of the original RARRRKKRT inhibitory scaffold produced selective and potent, nanomolar range, inhibitors of furin. These inhibitors did not interfere with the normal cellular function of furin because of the likely functional redundancy existing between furin and other proprotein convertases. These furin inhibitors, however, were highly potent in blocking the furin-dependent cell-surface processing of anthrax protective antigen-83 both in vitro and cell-based assays and in vivo. We conclude that the inhibitors we have designed have a promising potential as selective anthrax inhibitors, without affecting major cell functions.
弗林和相关的蛋白原转化酶切割前体蛋白中的多碱性基序 R-X-R/K/X-R,从而将潜伏的蛋白原转化为生物活性的蛋白质和肽。弗林存在于细胞内分泌途径和细胞表面。细胞内弗林在高尔基体和分泌小泡区室中处理其多个正常的细胞靶标,而细胞表面弗林似乎仅对某些致病蛋白的加工至关重要,重要的是炭疽。为了设计有效的、安全的和选择性的弗林抑制剂,我们评估了从禽流感 A H5N1 的扩展弗林切割序列建模的衍生肽抑制剂的效力和选择性。我们确定,原始 RARRRKKRT 抑制支架的 N-和 C-末端修饰产生了对弗林具有选择性和效力的纳摩尔范围的抑制剂。这些抑制剂不会干扰弗林的正常细胞功能,因为弗林和其他蛋白原转化酶之间可能存在功能冗余。这些弗林抑制剂在体外和基于细胞的测定以及体内都非常有效地阻止了炭疽保护性抗原 83 的弗林依赖性细胞表面加工。我们得出结论,我们设计的抑制剂具有作为选择性炭疽抑制剂的巨大潜力,而不会影响主要的细胞功能。