Oligosaccharides modulate the apoptotic activity of glycodelin.
GlycodelinA (GdA), a multifunctional glycoprotein secreted at high concentrations by the uterine endometrium during the early phases of pregnancy, carries glycan chains on asparagines at positions N28 and N63. GdA purified from amniotic fluid is known to be a suppressor of T-cell proliferation, an inducer of T-cell apoptosis, and an inhibitor of sperm-zona binding in contrast to its glycoform, glycodelinS (GdS), which is secreted by the seminal vesicles into the seminal plasma. The oligosaccharide chains of GdA terminate in sialic acid residues, whereas those of GdS are not sialylated but are heavily fucosylated. Our previous work has shown that the apoptogenic activity of GdA resides in the protein backbone, and we have also demonstrated the importance of sialylation for the manifestation of GdA-induced apoptosis. Recombinant glycodelin (Gd) expressed in the Sf21 insect cell line yielded an apoptotically active Gd; however, the same gene expressed in the insect cell line Tni produced apoptotically inactive Gd, as observed with the gene expressed in the Chinese hamster ovary (CHO) cell line and earlier in Pichia pastoris. Glycan analysis of the Tni and Sf21 cell line-expressed Gd proteins reveals differences in their glycan structures, which modulate the manifestation of apoptogenic activity of Gd. Through apoptotic assays carried out with the wild-type (WT) and glycosylation mutants of Gd expressed in Sf21 and Tni cells before and after mannosidase digestion, we conclude that the accessibility to the apoptogenic region of Gd is influenced by the size of the glycans.
Jayachandran
Rajesh
R
Department of Biochemistry, Indian Institute of Science, Balgalore 560012, India.
Radcliffe
Catherine M
CM
Royle
Louise
L
Harvey
David J
DJ
Dwek
Raymond A
RA
Rudd
Pauline M
PM
Karande
Anjali A
AA
D000818:Animals; D017209:Apoptosis; D001216:Asparagine / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D000071816:Glycodelin; D006023:Glycoproteins / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D006031:Glycosylation; D006801:Humans; D007313:Insecta; D019169:Jurkat Cells; D009154:Mutation; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D009844:Oligosaccharides / Q000494:pharmacology*; D011134:Polysaccharides / Q000378:metabolism; D011257:Pregnancy Proteins / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology
D000071816:Glycodelin; D006023:Glycoproteins; D009844:Oligosaccharides; C045246:PAEP protein, human; D011134:Polysaccharides; D011257:Pregnancy Proteins; D011994:Recombinant Proteins; D001216:Asparagine; D019158:N-Acetylneuraminic Acid