1976 Volume 26 Issue 2Supplement Pages 125-133
Phenol soluble proteinaceous substance (PSPS) was extracted from the whole cell of E. coli using the Westphal's phenol-water method.
The procedure was similar to that of the extraction of protodyne (Berger et al) excepting some difference in the starting materials of whole cell for extraction in the former and, in the latter, of the cytoplasma, previously isolated from the cell wall.
The results in regard to the chemical and biological characteristics of PSPS were as follow :
1) PSPS consisted mostly of proteins ; small amount of hexose and ribose, none of desoxyribose.
2) Using the Sephadex G-200 column chromatography the PSPS was separated into 3 fractions with the high absorption of 280 mμ.
3) Molecular weights were determined by Andrews' method as having 150, 000 for Fr I, 30, 000 for Fr II and 10, 000 for Fr III, respectively.
4) PSPS has an ability to induce the some degree of Shwartzman reaction in the rabbits, which was far less than that of endotoxin.
5) Homocytotropic antibody was produced by the PSPS incorporated in alum-gel as adjuvant; reached the maximum titer at 3 weeks; remained the high titer at 4 weeks after the immunization
In conclusion, PSPS looks like the protodyne as to the chemical characteristic except for the contamination with small amount of endotoxin, because of its relatively high contents of sugar and of inducible activity of the feeble Shwartzman reaction in rabbits in comparison with protodyne. From these results PSPS should be much more purifed by means of column chromatography to have an exclusively proteinaceous substance just like the protodyne.
