Edwards J A, Hines J F
Brain Res. 1982 May 13;239(2):507-17. doi: 10.1016/0006-8993(82)90525-x.
An in vitro preparation was used to determine if rapidly transported radiolabeled protein which is isolated with the myelin fraction of frog sciatic nerves is similar to rapidly transported radiolabeled protein released from sciatic nerves. Sciatic nerves along with their eighth and ninth dorsal root ganglia were placed in modified Warburg flasks. The center well of the flasks contained 100 microCi [3H]leucine. The preparation was incubated for 24 h, the sciatic nerve removed and the myelin isolated. The solution in which the sciatic nerve portion of the preparation incubated was removed, dialyzed and lyophilized. The radiolabeled proteins isolated from myelin and those released from the nerve were separated on 10% SDS polyacrylamide gels. Results of this separation revealed that a radiolabeled protein of 100,000 daltons was isolated with myelin and released from sciatic nerves. Incubating isolated myelin in 10 mM EDTA for 14-16 h brought about the release of a protein of 100,000 daltons.
采用体外制备方法来确定从蛙坐骨神经髓磷脂部分分离出的快速转运放射性标记蛋白是否与从坐骨神经释放的快速转运放射性标记蛋白相似。将坐骨神经及其第八和第九背根神经节置于改良的瓦尔堡烧瓶中。烧瓶的中心孔含有100微居里的[3H]亮氨酸。将制剂孵育24小时，取出坐骨神经并分离髓磷脂。将制剂中坐骨神经部分孵育的溶液取出，透析并冻干。从髓磷脂中分离出的放射性标记蛋白和从神经中释放出的放射性标记蛋白在10%十二烷基硫酸钠聚丙烯酰胺凝胶上进行分离。该分离结果显示，一种100,000道尔顿的放射性标记蛋白可从髓磷脂中分离出来并从坐骨神经中释放。将分离出的髓磷脂在10 mM乙二胺四乙酸中孵育14 - 16小时会导致一种100,000道尔顿的蛋白释放。
