Prem Das O, Henderson E J
J Cell Biol. 1983 Nov;97(5 Pt 1):1544-58. doi: 10.1083/jcb.97.5.1544.
Developmental changes in the plasma membrane proteins of Dictyostelium discoideum have been studied using metabolic labeling with [35S]methionine and two-dimensional electrophoresis. Pulse labeling for 1 h at the early interphase, late interphase, aggregation, and tip formation stages of development showed that the profile of newly synthesized plasma membrane proteins changed dramatically over this interval. Only 14% of the polypeptide species were synthesized at all four stages at detectable levels; 86% of the species changed over this developmental interval according to the criterion that they were synthesized at some but not all of the four stages tested. Long-term labeling during vegetative growth followed by initiation of development showed that the "steady-state" levels of the plasma membrane proteins changed very little over the same period. The only changes were in minor species (33% overall change). Similar analyses of whole cell proteins showed 27 and 20% change, respectively. Cell surface radioiodination revealed 52 external proteins in the plasma membrane. Comparison with the uniform methionine labeling results showed that these proteins were, with one notable exception, minor membrane components. In these external proteins, also, developmental changes were limited and were observed in the less abundant species. These results demonstrate the existence of two general classes of plasma membrane proteins. The first is a population of high-abundance proteins that are present in vegetative cells and are largely conserved through development. These possibly serve "housekeeping" functions common to all stages. The second class consists of low-abundance species that are expressed in a highly stage-specific manner and which presumably participate in developmentally important functions.
利用[35S]甲硫氨酸代谢标记和二维电泳技术,对盘基网柄菌质膜蛋白的发育变化进行了研究。在发育的早期间期、晚期间期、聚集期和尖端形成期进行1小时的脉冲标记,结果表明,在此期间新合成的质膜蛋白谱发生了显著变化。在所有四个阶段,只有14%的多肽种类能被检测到有合成;根据在四个测试阶段中的部分而非全部阶段有合成这一标准,86%的种类在这个发育期间发生了变化。在营养生长期间进行长期标记,然后开始发育,结果表明,质膜蛋白的“稳态”水平在同一时期变化很小。唯一的变化是在次要种类中(总体变化为33%)。对全细胞蛋白的类似分析分别显示变化为27%和20%。细胞表面放射性碘化显示质膜中有52种外部蛋白。与均匀甲硫氨酸标记结果比较表明,这些蛋白除了一个显著例外,都是次要的膜成分。在这些外部蛋白中,发育变化也很有限,且在丰度较低的种类中观察到。这些结果证明了质膜蛋白存在两大类。第一类是高丰度蛋白群体,存在于营养细胞中,在发育过程中基本保守。这些蛋白可能履行所有阶段共有的“看家”功能。第二类由低丰度种类组成,以高度阶段特异性的方式表达,推测参与重要的发育功能。