Elimination of potential sites of glycosylation fails to abrogate complement regulatory function of cell surface CD59.
CD59 is a glycosylphosphatidylinositol-anchored membrane glycoprotein that serves as the principle cellular inhibitor of the C5b-9 membrane attack complex (MAC) of human complement. Approximately 50% of the total apparent mass of CD59 is attributable to glycosylation of a single Asn (Asn18). The deduced amino acid sequences of CD59 homologues identified in Old and New World primates as well as in rat reveal that the motif for N-linked glycosylation at the residue corresponding to Asn18 of human CD59 is invariably conserved, despite considerable sequence divergence elsewhere in the protein. Such conservation suggests that the post-translational modification at Asn18 has importance for either expression or normal function of CD59 at the cell surface. In this study, we specifically examined how deletion or transposition of the site of N-linked glycosylation in the CD59 polypeptide affects its MAC inhibitory function. Our data demonstrate that the inhibitory potency of CD59 is unaffected when glycosylation is transposed from Asn18 to another site in the polypeptide. Furthermore, we show that CD59 retains normal MAC regulatory function when mutated to eliminate all potential sites for N-linked glycosylation. These data suggest that the MAC inhibitory function of CD59 is entirely provided by residues exposed at the surface of the core polypeptide and that this core structure is not influenced by glycosylation at Asn18.
Rother
R P
RP
Alexion Pharmaceuticals Inc, New Haven, Connecticut 06511, USA.
Zhao
J
J
Zhou
Q
Q
Sims
P J
PJ
D000595:Amino Acid Sequence; D001216:Asparagine / Q000737:chemistry; D018957:CD59 Antigens / Q000737:chemistry* / Q000502:physiology; D015938:Complement Membrane Attack Complex / Q000037:antagonists & inhibitors; D004252:DNA Mutational Analysis; D006023:Glycoproteins / Q000502:physiology; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
D018957:CD59 Antigens; D015938:Complement Membrane Attack Complex; D006023:Glycoproteins; D011994:Recombinant Proteins; D001216:Asparagine