Roberts K, Hills G J
J Cell Sci. 1976 Jun;21(1):59-71. doi: 10.1242/jcs.21.1.59.
Members of the Chlamydomonaceae, mostly single-celled green algae, have been shown to contain a crystalline glycoprotein cell wall component. Most of the species examined fall into a class of algae whose walls have an identical crystalline unit cell. Chlorogonium elongatum has been chosen as a representative of this class in order to investigate in more detail its cell wall structure. The alga has a spindleshaped cell wall which retains its asymmetric shape on isolation. Sections from walls fized in the presence of tannic acid clearly reveal a regular subunit monolayer, about 20 nm thick, within the wall. Sodium dodecylsulphate (SDS) polyacrylamide gel electrophoresis shows the presence of at least 2 major glycoprotein species in the wall. Negatively stained purified cell walls demonstrate the crystalline nature of the cell wall. Optical diffraction of bright-field images and direct electron diffraction both give clear diffraction patterns whose spacings extend out to 3 nm and fall on a reciprocal lattice whose vectors describe a 2-dimensional unit cell within the wall 21.5 nm X 7.0 nm and an included angle of 80 degrees. Lattice defects within the cell wall are revealed by both negative staining and surface replication. Through-focal series were used to choose images with the optimal degree of underfocus for image processing. Linear integration and optical filtering of such images gave essentially the same result. A similar image was also obtained by computing the autocorrelation function of the amplitudes in the electron-diffraction pattern and the optical-diffraction pattern of the in-focus image. On the basis of these data a 2-dimensional model of the crystalline cell wall layer is presented.
衣藻科的成员大多是单细胞绿藻,已被证明含有一种结晶糖蛋白细胞壁成分。大多数被检测的物种属于一类藻类,其细胞壁具有相同的结晶晶胞。为了更详细地研究其细胞壁结构,细长绿梭藻被选为该类别的代表。这种藻类有一个纺锤形的细胞壁,在分离时保持其不对称形状。在单宁酸存在下固定的细胞壁切片清楚地显示出壁内有一个规则的亚基单层,约20纳米厚。十二烷基硫酸钠(SDS)聚丙烯酰胺凝胶电泳显示壁中至少存在2种主要的糖蛋白。经负染色的纯化细胞壁证明了细胞壁的结晶性质。明场图像的光学衍射和直接电子衍射都给出了清晰的衍射图案,其间距延伸至3纳米,并落在一个倒易晶格上,其矢量描述了壁内一个21.5纳米×7.0纳米、夹角为80度的二维晶胞。通过负染色和表面复型都揭示了细胞壁内的晶格缺陷。使用透焦系列来选择具有最佳欠焦程度的图像进行图像处理。对这些图像进行线性积分和光学滤波得到的结果基本相同。通过计算聚焦图像的电子衍射图案和光学衍射图案中振幅的自相关函数,也获得了类似的图像。基于这些数据,提出了结晶细胞壁层的二维模型。