Axford Danny, Ji Xiaoyun, Stuart David I, Sutton Geoff
Diamond Light Source Ltd, Harwell Oxford, Didcot OX11 0DE, England.
Division of Structural Biology, The Welcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, England.
Acta Crystallogr D Biol Crystallogr. 2014 May;70(Pt 5):1435-41. doi: 10.1107/S1399004714004714. Epub 2014 Apr 30.
This work demonstrates that with the use of a microfocus synchrotron beam the structure of a novel viral polyhedrin could be successfully determined from microcrystals within cells, removing the preparatory step of sample isolation and maintaining a favourable biological environment. The data obtained are of high quality, comparable to that obtained from isolated crystals, and enabled a facile structure determination. A small but significant difference is observed between the unit-cell parameters and the mosaic spread of in cellulo and isolated crystals, suggesting that even these robust crystals are adversely affected by removal from the cell.
这项工作表明,使用微聚焦同步加速器光束,可以成功地从细胞内的微晶中确定一种新型病毒多角体蛋白的结构,省去了样品分离的制备步骤,并保持了良好的生物环境。获得的数据质量很高,与从分离晶体获得的数据相当,并且能够轻松地进行结构测定。在晶胞参数以及细胞内晶体和分离晶体的镶嵌角展度之间观察到一个微小但显著的差异,这表明即使是这些坚固的晶体从细胞中取出后也会受到不利影响。