Klein S M, Vernon L P
Biochim Biophys Acta. 1977 Mar 11;459(3):364-75. doi: 10.1016/0005-2728(77)90038-x.
The use of Triton X-100 to solubilize membrane fragments from Anabaena flos-aquae in conjunction with DEAE cellulose chromatography allows the separation of three green fractions. Fraction 1 is detergent-solubilized chlorophyll, and Fraction 2 contains one polypeptide in the 15 kdalton area. Fraction 3, which contains most of the chlorophyll and shows P-700 and photosystem I activity, shows by SDS gel electrophoresis varying polypeptide profiles which reflect the presence of four fundamental bands as well as varying amounts of other polypeptides which appear to be aggregates containing the 15 kdalton polypeptide. The four fundamental bands are designated Band I at 120, Band II at 52, Band III at 46, and Band IV at 15 kdaltons. Band I obtained using 0.1% SDS contains chlorophyll and P-700 associated with it. When this band is cut out and rerun, the 120 kdalton band is lost, but significant increases occur in the intensities of Bands II, III, and IV as well as other polypeptides in the 20-30 kdalton range. The use of 1% Triton X-100 coupled with sucrose density gradient centrifugation allows the separation of three green bands at 10, 25 and 40% sucrose. The 10% layer contains a major polypeptide which appears to be Band IV. The 25 and 40% layers show essentially similar polypeptide profiles, resembling Fraction 3 in this regard, except that the 40% layer shows a marked decrease in Band III. Treatment of the material layering at the 40% sucrose level with a higher (4%) concentration of Triton X-100 causes a loss (disaggregation) of the polypeptides occurring in the 60-80 kdalton region and in increase in the lower molecular weight polypeptides. Thus, aggregation of the lower molecular weight polypeptides accounts for the variability seen in the electrophoresis patterns. Possible relations of the principal polypeptides to the known photochemical functions in the original membrane are discussed.
使用 Triton X - 100 溶解水华鱼腥藻的膜片段并结合 DEAE 纤维素色谱法，可以分离出三个绿色组分。组分 1 是经去污剂溶解的叶绿素，组分 2 含有一个分子量在 15 千道尔顿区域的多肽。组分 3 含有大部分叶绿素，并显示出 P - 700 和光系统 I 的活性，通过 SDS 凝胶电泳显示出不同的多肽图谱，反映出四条基本条带的存在以及不同数量的其他多肽，这些多肽似乎是含有 15 千道尔顿多肽的聚集体。这四条基本条带分别被指定为：120 千道尔顿的条带 I、52 千道尔顿的条带 II、46 千道尔顿的条带 III 和 15 千道尔顿的条带 IV。使用 0.1% SDS 获得的条带 I 含有与之相关的叶绿素和 P - 700。当切下这条带并重新进行电泳时，120 千道尔顿的条带消失，但条带 II、III 和 IV 以及 20 - 30 千道尔顿范围内的其他多肽的强度显著增加。使用 1% Triton X - 100 并结合蔗糖密度梯度离心，可以在 10%、25%和 40%蔗糖浓度下分离出三个绿色条带。10%蔗糖层含有一种主要多肽，似乎是条带 IV。25%和 40%蔗糖层显示出基本相似的多肽图谱，在这方面与组分 3 相似，只是 40%蔗糖层的条带 III 明显减少。用较高浓度（4%）的 Triton X - 100 处理在 40%蔗糖水平分层的材料，会导致 60 - 80 千道尔顿区域出现的多肽损失（解聚），并使低分子量多肽增加。因此，低分子量多肽的聚集解释了电泳图谱中观察到的变异性。文中还讨论了主要多肽与原始膜中已知光化学功能的可能关系。
