TY - JOUR
T1 - A necessary and sufficient determinant for protein-selective glycosylation in vivo
AU - Miller, Erin
AU - Fiete, Dorothy
AU - Blake, Nicquet M.J.
AU - Beranek, Mary
AU - Oates, Edward L.
AU - Mi, Yiling
AU - Roseman, Daniel S.
AU - Baenziger, Jacques U.
PY - 2008/1/25
Y1 - 2008/1/25
N2 - A limited number of glycoproteins including luteinizing hormone and carbonic anhydrase-VI (CA6) bear N-linked oligosaccharides that are modified with β1,4-linked N-acetylgalactosamine (GalNAc). The selective addition of GalNAc to these glycoproteins requires that the β1,4-N- acetylgalactosaminyl-transferase (βGT) recognize both the oligosaccharide acceptor and a peptide recognition determinant on the substrate glycoprotein. We report here that two recently cloned βGTs, βGT3 and βGT4, that are able to transfer GalNAc to GlcNAc in β1,4-linkage display the necessary glycoprotein specificity in vivo. Both βGTs transfer GalNAc to N-linked oligosaccharides on the luteinizing hormone α subunit and CA6 but not to those on transferrin (Trf). A single peptide recognition determinant encoded in the carboxyl-terminal 19-amino acid sequence of bovine CA6 mediates transfer of GalNAc to each of its two N-linked oligosaccharides. The addition of this 19-amino acid sequence to the carboxyl terminus of Trf confers full acceptor activity onto Trf for both βGT3 and βGT4 in vivo. The complete 19-amino acid sequence is required for optimal GalNAc addition in vivo, indicating that the peptide sequence is both necessary and sufficient for recognition by βGT3 and βGT4.
AB - A limited number of glycoproteins including luteinizing hormone and carbonic anhydrase-VI (CA6) bear N-linked oligosaccharides that are modified with β1,4-linked N-acetylgalactosamine (GalNAc). The selective addition of GalNAc to these glycoproteins requires that the β1,4-N- acetylgalactosaminyl-transferase (βGT) recognize both the oligosaccharide acceptor and a peptide recognition determinant on the substrate glycoprotein. We report here that two recently cloned βGTs, βGT3 and βGT4, that are able to transfer GalNAc to GlcNAc in β1,4-linkage display the necessary glycoprotein specificity in vivo. Both βGTs transfer GalNAc to N-linked oligosaccharides on the luteinizing hormone α subunit and CA6 but not to those on transferrin (Trf). A single peptide recognition determinant encoded in the carboxyl-terminal 19-amino acid sequence of bovine CA6 mediates transfer of GalNAc to each of its two N-linked oligosaccharides. The addition of this 19-amino acid sequence to the carboxyl terminus of Trf confers full acceptor activity onto Trf for both βGT3 and βGT4 in vivo. The complete 19-amino acid sequence is required for optimal GalNAc addition in vivo, indicating that the peptide sequence is both necessary and sufficient for recognition by βGT3 and βGT4.
UR - http://www.scopus.com/inward/record.url?scp=38349152485&partnerID=8YFLogxK
U2 - 10.1074/jbc.M708160200
DO - 10.1074/jbc.M708160200
M3 - Article
C2 - 18048353
AN - SCOPUS:38349152485
VL - 283
SP - 1985
EP - 1991
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 4
ER -