Department of Plant Physiology, University of Amsterdam, Kruislaan 318, Amsterdam, The Netherlands.
Plant Physiol. 1985 Nov;79(3):740-5. doi: 10.1104/pp.79.3.740.
Gametes of the unicellular green alga Chlamydomonas eugametos agglutinate via their flagella. The mating type plus agglutination factor was solubilized by relatively mild treatments such as a short pH shock or an osmotic shock indicating that it is an extrinsic membrane component. It was also extracted in the nonionic detergent Triton X-100. A simple two-step procedure consisting of gel filtration over Sepharose 4B-cross-linked followed by anion exchange chromatography of the void volume yielded an electrophoretically pure preparation of a single high molecular weight glycoprotein. The agglutination factor sedimented as a 9.3 S particle (assuming a density of 1.50) in sucrose gradients. This low value, compared with the high apparent molecular weight seen during gel filtration and electrophoresis, suggests that the agglutination factor is a rod-like molecule. This was confirmed by viewing rotary-shadowed preparations in the electron microscope. A population of long slender molecules was revealed (328 +/- 20 nanometers), many of which had a knob at one end and a flexible region about one fourth of the length from the other end.
单细胞绿藻衣藻的配子通过鞭毛凝集。交配型加凝集因子可通过相对温和的处理(如短时间 pH 冲击或渗透压冲击)溶解,表明它是一种外膜成分。它也可以用非离子去污剂 Triton X-100 提取。一个简单的两步程序,包括 Sepharose 4B 交联凝胶过滤,然后是空隙体积的阴离子交换层析,可得到电泳纯的单一高分子量糖蛋白制剂。凝集因子在蔗糖梯度中沉降为 9.3 S 颗粒(假设密度为 1.50)。与凝胶过滤和电泳中观察到的高表观分子量相比,这个低值表明凝集因子是一种棒状分子。这通过在电子显微镜下观察旋转阴影制备物得到证实。揭示了一群长而细的分子(328 +/- 20 纳米),其中许多分子在一端有一个旋钮,在另一端的四分之一长度处有一个柔性区域。