Trang Dai Thi Xuan, Huy Nguyen Tien, Uyen Dinh Thanh, Sasai Motohiro, Shiono Takeshi, Harada Shigeharu, Kamei Kaeko
Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
Anal Biochem. 2006 Feb 15;349(2):292-6. doi: 10.1016/j.ab.2005.11.022. Epub 2005 Dec 6.
Measurement of heme crystallization provides a tool for screening new antimalarial drugs. Current assays for heme crystallization have employed initiators such as thermo, histidine-rich proteins, and lipids extracted from parasites and infected plasma. These initiators are unnatural or require laborious steps to prepare. In this study, we used a commercially available lipid, lecithin, a kind of phospholipid containing about 50% unsaturated fatty acids, as an initiator for heme crystal (beta-hematin) formation. We demonstrated that the inhibition of lecithin-based beta-hematin formation by antimalarial drugs is highly correlated with the preformed beta-hematin-based method. In addition, the lecithin-based assay is sensitive and convenient for large-scale screening of new novel antimalarials. We also indicated that dimethyl sulfoxide is an ideal solvent for preparation of heme stock solution, which is stable and can be used for 1 month.
血红素结晶的测量为筛选新型抗疟药物提供了一种工具。目前用于血红素结晶的检测方法采用了诸如热、富含组氨酸的蛋白质以及从寄生虫和感染血浆中提取的脂质等引发剂。这些引发剂是非天然的，或者需要繁琐的步骤来制备。在本研究中，我们使用了一种市售脂质——卵磷脂，一种含有约50%不饱和脂肪酸的磷脂，作为血红素晶体（β-血红素）形成的引发剂。我们证明，抗疟药物对基于卵磷脂的β-血红素形成的抑制作用与基于预先形成的β-血红素的方法高度相关。此外，基于卵磷脂的检测方法对于大规模筛选新型抗疟药物而言灵敏且便捷。我们还表明，二甲基亚砜是制备血红素储备溶液的理想溶剂，该储备溶液稳定且可使用1个月。
