Klausner R D, Kumar N, Weinstein J N, Blumenthal R, Flavin M
J Biol Chem. 1981 Jun 10;256(11):5879-85.
Purified soluble tubulin will interact with highly sonicated small unilamellar vesicles of dipalmitoyl phosphatidylcholine. This results in the leakage of a fluorescent dye, carboxyfluorescein, from the internal aqueous space of the vesicles at the lipid phase transition. The amount of dye leaked from the vesicles is determined by the tubulin to vesicle ratio and partial dye leakage reflects the uniform leakage of carboxyfluorescein from all of the vesicles. Once tubulin interacts with dipalmitoyl phosphatidylcholine vesicles at the phase transition, it is unavailable to interact with additional vesicles. This interaction represents the formation of stable tubulin-vesicle recombinants as shown by density gradient centrifugation. Such recombinants are only formed by interacting the protein with the vesicles at the lipid phase transition temperature. The interaction is insensitive to ionic strength (0.001 to 4 M NaCl) and the recombinants remain stable for days.
纯化的可溶性微管蛋白会与高度超声处理的二棕榈酰磷脂酰胆碱小单层囊泡相互作用。这会导致荧光染料羧基荧光素在脂质相变时从囊泡的内部水相空间泄漏。从囊泡泄漏的染料量由微管蛋白与囊泡的比例决定，部分染料泄漏反映了羧基荧光素从所有囊泡中均匀泄漏。一旦微管蛋白在相变时与二棕榈酰磷脂酰胆碱囊泡相互作用，它就无法再与其他囊泡相互作用。如密度梯度离心所示，这种相互作用代表了稳定的微管蛋白 - 囊泡重组体的形成。这种重组体仅在脂质相变温度下通过使蛋白质与囊泡相互作用而形成。这种相互作用对离子强度（0.001至4 M NaCl）不敏感，并且重组体可保持稳定数天。
