M. Flotenmeyer et al., Hydrophobic and hydrophilic radio-iodination, crosslinking, and differential extraction of cell surface proteins in Paramecium tetraurelia cells, J MEMBR BIO, 172(1), 1999, pp. 77-88
We combined widely different biochemical methods to analyze proteins of the
cell surface of P. tetraurelia since so far one can isolate only a subfrac
tion of cell membrane vesicles enriched in the GPI-anchored surface antigen
s ("immoblization" or "i-AGs"). We also found that i-AGs may undergo partia
l degradation by endogenous proteases. Genuine intrinsic membrane proteins
were recognized particularly with lipophilic 5-[I-125]-iodonaphthalene-1-az
ide (INA) labeling which reportedly "sees" integral proteins and cytoplasmi
c cell membrane-associated proteins. With INA (+DTT), bands of less than or
equal to 55 kDa were similar as with hydrophilic iodogen (+DTT), but inste
ad of large size bands including i-AGs, a group of 122, 104 and 94 kDa appe
ared. Several bands of the non i-AG type are compatible with integral (poss
ibly oligomeric) or associated proteins of the cell membrane of established
molecular identity, as we discuss. In summary, we can discriminate between
i-AGs and some functionally important minor cell membrane components. Our
methodical approach might be relevant also for an analysis of some related
protozoan parasites.