Dürr M, Boller T, Wiemken A
Arch Microbiol. 1975 Nov 7;105(3):319-27. doi: 10.1007/BF00447152.
The polybasic macromolecules DEAE-dextran (diethylaminoethyl-dextran, molecular weight 500000) and poly-DL-lysine (molecular weight 30000-70000) were absorbed with a high affinity by spheroplasts of Candida utilis and subsequently, induced lysis. The extent of lysis of spheroplasts and of the liberated vacuoles was studied under various conditions using alpha-glucosidase activity and soluble arginine as cytoplasmic and vacuolar markers, respectively. Adsorption of polybases was rapidly completed even at 0 degrees C; however, with small doses, lysis was poor at 0-12 degrees C and extensive at temperatures above 12 degrees C. This permitted the completion of adsorption before initiating lysis. The purified vacuoles were also sensitive to polybases though less so than the spheroplasts; however, after lysis of spheroplasts the liberated vacuoles were well protected against the action of polybases. A treatment with polybases which disrupted more than 99% of the spheroplasts left at least 70% of the vacuoles intact. Potassium chloride in high concentrations and calcium chloride in low concentrations inhibited polybase induced lysis of spheroplasts by preventing or even reversing the polybase adsorption. A polyacidic macromolecule, dextran sulfate, could prevent but not reverse the adsorption of polybase and subsequent lysis. Metabolic inhibitors reduced the susceptibility of spheroplasts to polybase induced lysis. Vacuoles isolated from polybase lysed spheroplasts still contained large pools of soluble amino acids, and their ability to transport arginine specifically is a further indication of their functional integrity.
多碱性大分子二乙氨基乙基葡聚糖（DEAE - 葡聚糖，分子量500000）和聚 - DL - 赖氨酸（分子量30000 - 70000）被产朊假丝酵母原生质体以高亲和力吸收，随后诱导裂解。在各种条件下，分别使用α - 葡萄糖苷酶活性和可溶性精氨酸作为细胞质和液泡标记物，研究原生质体和释放出的液泡的裂解程度。即使在0℃，多碱的吸附也能迅速完成；然而，使用小剂量时，在0 - 12℃裂解效果不佳，而在高于12℃时裂解广泛。这使得在开始裂解之前能够完成吸附。纯化的液泡对多碱也敏感，尽管不如原生质体敏感；然而，原生质体裂解后释放出的液泡对多碱的作用有很好的保护。用多碱处理破坏了超过99%的原生质体，但至少70%的液泡保持完整。高浓度的氯化钾和低浓度的氯化钙通过阻止甚至逆转多碱吸附来抑制多碱诱导的原生质体裂解。一种多酸性大分子硫酸葡聚糖可以阻止但不能逆转多碱的吸附及随后的裂解。代谢抑制剂降低了原生质体对多碱诱导裂解的敏感性。从多碱裂解的原生质体中分离出的液泡仍然含有大量可溶性氨基酸库，它们特异性转运精氨酸的能力进一步表明了其功能完整性。
