Darvish-Damavandi Mahnaz, Ho Han Kiat, Kang Tse Siang
Department of Pharmacy, Faculty of Science, National University of Singapore, Singapore.
Mol Genet Metab Rep. 2016 Jul 27;8:51-60. doi: 10.1016/j.ymgmr.2016.06.009. eCollection 2016 Sep.
Propionic acidemia (PA) is a life-threatening disease caused by the deficiency of a mitochondrial biotin-dependent enzyme known as propionyl coenzyme-A carboxylase (PCC). This enzyme is responsible for degrading the metabolic intermediate, propionyl coenzyme-A (PP-CoA), derived from multiple metabolic pathways. Currently, except for drastic surgical and dietary intervention that can only provide partial symptomatic relief, no other form of therapeutic option is available for this genetic disorder. Here, we examine a novel approach in protein delivery by specifically targeting and localizing our protein candidate of interest into the mitochondrial matrix of the cells. In order to test this concept of delivery, we have utilized cell penetrating peptides (CPPs) and mitochondria targeting sequences (MTS) to form specific fusion PCC protein, capable of translocating and localizing across cell membranes. In vitro delivery of our candidate fusion proteins, evaluated by confocal images and enzymatic activity assay, indicated effectiveness of this strategy. Therefore, it holds immense potential in creating a new paradigm in site-specific protein delivery and enzyme replacement therapeutic for PA.
丙酸血症（PA）是一种由线粒体生物素依赖性酶——丙酰辅酶A羧化酶（PCC）缺乏引起的危及生命的疾病。该酶负责降解来自多种代谢途径的代谢中间体丙酰辅酶A（PP-CoA）。目前，除了只能提供部分症状缓解的激进手术和饮食干预外，这种遗传性疾病没有其他治疗选择。在这里，我们研究了一种蛋白质递送的新方法，即将我们感兴趣的蛋白质候选物特异性靶向并定位到细胞的线粒体基质中。为了测试这种递送概念，我们利用细胞穿透肽（CPP）和线粒体靶向序列（MTS）形成特定的融合PCC蛋白，该蛋白能够跨细胞膜转运和定位。通过共聚焦图像和酶活性测定评估我们的候选融合蛋白的体外递送，表明了该策略的有效性。因此，它在为PA创造位点特异性蛋白质递送和酶替代治疗的新范例方面具有巨大潜力。
