Broo K, Wei J, Marshall D, Brown F, Smith T J, Johnson J E, Schneemann A, Siuzdak G
The Scripps Research Institute, Center for Mass Spectrometry and Department of Molecular Biology, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2274-7. doi: 10.1073/pnas.051598298. Epub 2001 Feb 13.
Mass spectrometry and fluorescent probes have provided direct evidence that alkylating agents permeate the protein capsid of naked viruses and chemically inactivate the nucleic acid. N-acetyl-aziridine and a fluorescent alkylating agent, dansyl sulfonate aziridine, inactivated three different viruses, flock house virus, human rhinovirus-14, and foot and mouth disease virus. Mass spectral studies as well as fluorescent probes showed that alkylation of the genome was the mechanism of inactivation. Because particle integrity was not affected by selective alkylation (as shown by electron microscopy and sucrose gradient experiments), it was reasoned that the dynamic nature of the viral capsid acts as a conduit to the interior of the particle. Potential applications include fluorescent labeling for imaging viral genomes in living cells, the sterilization of blood products, vaccine development, and viral inactivation in vivo.
质谱分析和荧光探针已提供直接证据表明,烷基化剂可穿透裸露病毒的蛋白质衣壳并使核酸化学失活。N-乙酰氮丙啶和一种荧光烷基化剂——丹磺酰磺酸氮丙啶,可使三种不同的病毒失活,即鸡瘟病毒、人鼻病毒-14和口蹄疫病毒。质谱研究以及荧光探针表明,基因组的烷基化是失活机制。由于选择性烷基化未影响颗粒完整性(如电子显微镜和蔗糖梯度实验所示),因此据推测病毒衣壳的动态特性充当了通向颗粒内部的管道。潜在应用包括用于对活细胞中的病毒基因组进行成像的荧光标记、血液制品的灭菌、疫苗开发以及体内病毒失活。