Krebs K E, Zagon I S, Goodman S R
Brain Res Bull. 1986 Aug;17(2):237-41. doi: 10.1016/0361-9230(86)90120-6.
We have developed a one chromatographic step isolation protocol for the neuron specific protein synapsin I. This procedure results in a yield of 80 micrograms/g brain, which is ten fold better than the highest yield yet reported for this protein. The authenticity of the synapsin I isolated by this procedure is demonstrated by comigration with authentic synapsin I on SDS-polyacrylamide gels, crossreactivity with antibody specific against synapsin I, and nearly identical two dimensional chrymotryptic iodopeptide maps of authentic synapsin I and the protein purified by this protocol. Synapsin I isolated by this procedure retains its functional properties, demonstrated by the ability of synapsin I to stimulate the formation of a brain spectrin(240/235)/synapsin I/F-actin ternary complex as determined by a low shear falling ball viscometry assay. This novel protocol therefore has the advantage of being a rapid, high yield procedure that retains the functional properties of synapsin I.
我们已经开发出一种用于分离神经元特异性蛋白突触素I的单步色谱分离方案。该方法的产量为80微克/克脑组织，比该蛋白迄今报道的最高产量高出十倍。通过在SDS-聚丙烯酰胺凝胶上与真实的突触素I共迁移、与针对突触素I的特异性抗体发生交叉反应以及真实突触素I与通过该方案纯化的蛋白几乎相同的二维胰凝乳蛋白酶解碘肽图谱，证明了通过该方法分离的突触素I的真实性。通过该方法分离的突触素I保留了其功能特性，如通过低剪切落球粘度测定法所确定的，突触素I能够刺激脑血影蛋白（240/235）/突触素I/F-肌动蛋白三元复合物的形成。因此，这种新颖的方案具有快速、高产且保留突触素I功能特性的优点。
