Goodenough U W, Gebhart B, Mecham R P, Heuser J E
J Cell Biol. 1986 Aug;103(2):405-17. doi: 10.1083/jcb.103.2.405.
Two of the three major outer layers of the Chlamydomonas reinhardtii cell wall (W6 and W4) can be solubilized from living cells with sodium perchlorate or other chaotropes and will repolymerize in vitro to form milligram amounts of wall crystals. Conditions for optimal crystalization are presented, and conditions that fail to induce polymerization are exploited to maintain monomers in aqueous solution for ion-exchange chromatography. The four major glycoproteins of the complex (GP1, 1.5, 2, and 3) have in this way been purified to apparent homogeneity and have been characterized morphologically by transmission electron microscopy using the quick-freeze, deep-etch technique and by amino acid composition. Three of the four are hydroxyproline-rich species that copolymerize to form the W6 layer. The fourth (GP1.5) is a glycine-rich species that binds to the interior of the in vitro crystal; it is apparently equivalent to the granules within the W4 layer in situ.
莱茵衣藻细胞壁的三个主要外层中的两个（W6和W4）可以用高氯酸钠或其他离液剂从活细胞中溶解出来，并在体外重新聚合形成毫克量的壁晶体。本文给出了最佳结晶条件，并利用未能诱导聚合的条件将单体保持在水溶液中用于离子交换色谱。通过这种方式，复合物的四种主要糖蛋白（GP1、1.5、2和3）已被纯化至表观均一，并通过快速冷冻、深度蚀刻技术的透射电子显微镜和氨基酸组成进行了形态学表征。四种中的三种是富含羟脯氨酸的物种，它们共聚形成W6层。第四种（GP1.5）是富含甘氨酸的物种，它结合到体外晶体的内部；它显然等同于原位W4层内的颗粒。
