Sialylmotifs of sialyltransferases.
The sialyl moiety of sialylated glycoconjugates expressed on the cell surface are increasingly recognized as the key determinants of various biological recognition events. The transfer of sialic acid to these glycoconjugates are catalyzed by sialyltransferases, a group of 15 or more Golgi enzymes. Cloning of three sialyltransferases from this laboratory, indicated for the first time, that these enzymes are type II membrane proteins and share the topological features common to other glycosyltransferases. However, unlike the other members of the glycosyltransferase family, these enzymes showed the presence of two conserve protein domains, termed 'sialylmotifs'. This unique feature was subsequently found to be present in all the sialyltransferases cloned to-date. The larger 'L-sialylmotif' consisting of 48-49 amino acids is present in the middle of the luminal catalytic domain and has, eight invariant residues, while the 'S-sialylmotif' present closer to the C-terminal end of the enzyme has two invariants among a stretch of 23 amino acids. The other not-so-invariant amino acids are also conserved and their replacement is limited. The functional role of these two sialylmotifs were investigated by single-point site-directed mutagenesis using Gal beta 1, 4GlcNAc alpha 2,6-sialyltransferase (ST6Gal I) as a model. Detailed kinetic analysis of the mutants indicated that the 'L-sialylmotif' contributes to the binding of the common donor substrate CMP-NeuAc, while the 'S-sialylmotif' contributes to the binding of both the donor and acceptor substrates.
Datta
A K
AK
Cytel Corporation, Scripps Research Institute, San Diego, California 92121, USA. arun_datta@cytelcorp.com
Paulson
J C
JC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017386:Sequence Homology, Amino Acid; D012799:Sialyltransferases / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; D012799:Sialyltransferases