Rat aglycotransferrin and human monoglycotransferrin: production and metabolic properties.
Rat transferrin (rTf), containing one complex glycan, and human transferrin (hTf), containing two complex glycans, were treated with peptide:N-glycosidase F (PNGase) under nondenaturing conditions. Apo rTf with a nonfucosylated standard biantennary glycan, but not its diferric counterpart, yielded satisfactory amounts (approximately 55% in 7 h) of aglyco Tf (AgTf). The presence of a chitobiose core fucose reduced yields to approximately 30%, whereas an additional NeuAc on the GlcNAc in the Man(alpha 1-3) branch had no adverse effect. Triton X-100 impaired deglycosylation. The main product (approximately 65%) obtained from apo hTf was monoglyco Tf (MgTf). Analysis of the cyanogen bromide fragments of MgTf revealed that PNGase did not discriminate between the two glycosylation sites of hTf. A negligible portion (2-4%) of AgTf, that was also obtained during the reaction, probably resulted from PNGase action on denatured hTf molecules. Modified Tfs were separated by affinity chromatography, radiolabeled, mixed with another preparation of interest, and injected in rats. Total-body radiation measurements showed that the half-life of rat AgTf was 19-20% shorter than that of rTf but 9% longer than that of asialo Tf. This suggests that close to 76% of the change in the degradative rate observed after desialylating rTf is referable to charge loss rather than the exposure of Gal residues. Human MgTf was catabolized by rats 8-9% faster than the parent protein, while human AgTf behaved in vivo like a denatured protein. It is concluded that sialyl carboxyls are a codeterminant of the normal lifetime of transferrins.
Regoeczi
E
E
Department of Pathology, McMaster University Health Sciences Centre, Hamilton, Ontario, Canada.
Bolyos
M
M
Chindemi
P A
PA
D000818:Animals; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification / Q000493:pharmacokinetics*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D051381:Rats; D011994:Recombinant Proteins; D014168:Transferrin / Q000302:isolation & purification / Q000493:pharmacokinetics*
D006023:Glycoproteins; D011994:Recombinant Proteins; D014168:Transferrin; D006026:Glycoside Hydrolases