Fisher M., Pick U., Zamir A.
Biochemistry Department, Weizmann Institute of Science, Rehovot 76100, Israel.
Plant Physiol. 1994 Dec;106(4):1359-1365. doi: 10.1104/pp.106.4.1359.
The halotolerant alga Dunaliella salina grows in saline conditions as varied as 0.5 and 5 M NaCl, maintaining throughout this range a low intracellular ion concentration. To discover factors potentially involved in ionic homeostasis, we grew cells in media with different salinities or osmolarities and compared their protein profiles. The comparisons indicated that the amount of a 60-kD protein, p60, greatly increased with an increase in salinity and was moderately enhanced when NaCl was substituted with iso-osmotic glycerol. Cells transferred from low to high NaCl or from high glycerol to iso-osmotic NaCl media transiently ceased to grow, and resumption of growth coincided approximately with an increase in p60. The protein, extracted from a plasma membrane fraction, was purified to homogeneity. Anti-p60 antibodies cross-reacted with a 60-kD protein in Dunaliella bardawil. Immunoelectron microscopy of D. salina cell sections indicated that p60 was exclusively located in the plasma membrane. Its induction by salt, the correlation between its accumulation and growth resumption in high concentrations of salt, and its plasma membrane localization suggest the possibility that p60 could play a role in ionic homeostasis in conditions of high salinity, although different types of function could also be considered.
耐盐藻类杜氏盐藻能在盐浓度为0.5至5M NaCl的多种盐环境中生长,在此整个范围内其细胞内离子浓度都维持在较低水平。为了发现可能参与离子稳态的因素,我们将细胞培养在具有不同盐度或渗透压的培养基中,并比较它们的蛋白质谱。比较结果表明,一种60kD蛋白质p60的量随着盐度的增加而大幅增加,当用等渗甘油替代NaCl时,其含量也适度增加。从低盐度转移至高盐度NaCl环境或从高甘油环境转移至等渗NaCl培养基的细胞会短暂停止生长,生长的恢复大致与p60的增加同时发生。从质膜组分中提取的该蛋白质被纯化至同质。抗p60抗体与巴氏杜氏盐藻中的一种60kD蛋白质发生交叉反应。杜氏盐藻细胞切片的免疫电子显微镜观察表明,p60仅位于质膜中。它受盐诱导,其积累与在高盐浓度下生长恢复之间的相关性,以及其质膜定位表明,p60有可能在高盐度条件下的离子稳态中发挥作用,不过也可以考虑其具有不同类型的功能。