Togashi Shuntaro, Takahashi Kyosuke, Tamura Arisa, Toyota Ikumi, Hatakeyama Shiori, Komatsuda Atsushi, Kudo Ikuru, Sasaki Kudoh Erina, Okamoto Tomoya, Haga Asami, Miyamoto Asuka, Grave Ewa, Sugawara Taku, Shimizu Hiroaki, Itoh Hideaki
Department of Neurosurgery, Akita University Graduate School of Medicine, Akita 010-8543, Japan.
Department of Life Science, Akita University Graduate School of Engineering Science, Akita 010-8502, Japan.
J Biochem. 2017 Jul 1;162(1):27-36. doi: 10.1093/jb/mvw104.
Colistin is an antimicrobial cationic peptide that belongs to the polymyxin family. Colistin was clinically used for the treatment of gram-negative infections but fell out of favour because of its significant side effects including neurotoxicity and nephrotoxicity. More recently, colistin has been regarded as one of the important options for nosocomial infections caused by multidrug resistant bacteria. Mechanisms of both the side effect onset of the drug and the side effect reduction are yet to be elucidated. In this study, we identified the specific binding protein of colistin using an affinity column chromatography. Colistin binds to the molecular chaperone HSP90. Although colistin slightly suppressed the chaperone activity of HSP90, there are no effects on the ATPase activity for a low concentration of colistin. Interestingly, colistin-induced aggregation of HSP90 via the N-domain. As for the cell viability of the SHSY5Y cell, the cell viability decreased to approximately 80% by the colistin 300 μM. However, the cell viability recovered to approximately 100% by adding ATP dosage. The same result was obtained by dot blot assay using anti-HSP90 antibody. Our results may help to understand the side effect mechanism of colistin.
黏菌素是一种属于多黏菌素家族的抗菌阳离子肽。黏菌素曾在临床上用于治疗革兰氏阴性菌感染，但由于其显著的副作用，包括神经毒性和肾毒性，而不再受青睐。最近，黏菌素被视为治疗多重耐药菌引起的医院感染的重要选择之一。该药物副作用发生和副作用减轻的机制尚待阐明。在本研究中，我们使用亲和柱色谱法鉴定了黏菌素的特异性结合蛋白。黏菌素与分子伴侣HSP90结合。尽管黏菌素轻微抑制了HSP90的伴侣活性，但低浓度的黏菌素对ATP酶活性没有影响。有趣的是，黏菌素通过N结构域诱导HSP90聚集。至于SHSY5Y细胞的细胞活力，300μM的黏菌素使细胞活力降至约80%。然而，通过添加ATP剂量，细胞活力恢复到约100%。使用抗HSP90抗体的斑点印迹分析也得到了相同的结果。我们的结果可能有助于理解黏菌素的副作用机制。
