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A soluble extract from purified cell walls of C. reinhardii has been separated by gel filtration into three fractions which together account for 94% of the cell wall. The major fraction (accounting for 70% of the extract) is a glycoprotein, with a molecular wt. in sodium perchlorate of 298,000, which can be split into 4 electrophoretically distinct species. It contains 35% protein with high levels of hydroxyproline, arabinose and galactose, and is capable of self assembly into crystalline structures identical to those found within the cell wall. The second fraction (25% of the extract) is a similar glycoprotein, but contains 24% protein, a higher proportion of mannose, and is incapable of self assembly. The third fraction (3–6% of the extract) is shown to be an adsorbed impurity from the growth medium used.

We have investigated the in vitro reassembly of the salt soluble, hydroxyproline rich, glycoproteins from the cell wall of Chlamydomonas reinhardii, into structured cell wall fragments. We have devised an assay which has been used to follow the reassembly of the unfractionated and fractionated (2BI and 2BII) cell wall glycoproteins. Reassembly has a pH optimum of 5, a temperature optimum of 20°C, and the final size of the reassembled fragments appears to be promoted by the minor component 2BI. Periodate oxidation experiments show that sugar residues, in particular mannose, are important for accurate reassembly. Using electron microscopy, the structure of the reassembled products has been elucidated, as have intermediate stages in the reassembly process.