Glycosylphosphatidylinositol anchors of membrane glycoproteins are binding determinants for the channel-forming toxin aerolysin.
Cells that are sensitive to the channel-forming toxin aerolysin contain surface glycoproteins that bind the toxin with high affinity. Here we show that a common feature of aerolysin receptors is the presence of a glycosylphosphatidylinositol anchor, and we present evidence that the anchor itself is an essential part of the toxin binding determinant. The glycosylphosphatidylinositol (GPI)-anchored T-lymphocyte protein Thy-1 is an example of a protein that acts as an aerolysin receptor. This protein retained its ability to bind aerolysin when it was expressed in Chinese hamster ovary cells, but could not bind the toxin when expressed in Escherichia coli, where the GPI anchor is absent. An unrelated GPI-anchored protein, the variant surface glycoprotein of trypanosomes, was shown to bind aerolysin with similar affinity to Thy-1, and this binding ability was significantly reduced when the anchor was removed chemically. Cathepsin D, a protein with no affinity for aerolysin, was converted to an aerolysin binding form when it was expressed as a GPI-anchored hybrid in COS cells. Not all GPI-anchored proteins bind aerolysin. In some cases this may be due to differences in the structure of the anchor itself. Thus the GPI-anchored proteins procyclin of Trypanosoma congolense and gp63 of Leishmania major did not bind aerolysin, but when gp63 was expressed with a mammalian GPI anchor in Chinese hamster ovary cells, it bound the toxin.
Diep
D B
DB
Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
Nelson
K L
KL
Raja
S M
SM
Pleshak
E N
EN
Buckley
J T
JT
D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D001921:Brain / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000378:metabolism*; D007473:Ion Channels / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D052899:Pore Forming Cytotoxic Proteins; D011485:Protein Binding; D013601:T-Lymphocytes / Q000737:chemistry / Q000378:metabolism; D018800:Thy-1 Antigens / Q000032:analysis
D001427:Bacterial Toxins; D017261:Glycosylphosphatidylinositols; D006460:Hemolysin Proteins; D007473:Ion Channels; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D018800:Thy-1 Antigens; C107096:proaerolysin; C032676:aerolysin