N. Kurosawa et al., MOLECULAR-CLONING AND EXPRESSION OF CHICK-EMBRYO GAL-BETA-1,4GLCNAC-ALPHA-2,6-SIALYLTRANSFERASE - COMPARISON WITH THE MAMMALIAN ENZYME, European journal of biochemistry, 219(1-2), 1994, pp. 375-381
DNA clones encoding beta-galactoside alpha 2,6-sialyltransferase have
been isolated from chick embryonic cDNA libraries using sequence infor
mation obtained from the conserved amino acid sequence of the previous
ly cloned enzymes. The cDNA sequence revealed an open reading frame co
ding for 413 amino acids, and the deduced amino acid sequence showed 5
7.6% identity with the sequence of rat liver Gal beta 1,4GlcNAc alpha
2,6-sialyltransferase. The primary structure of this enzyme suggested
a putative domain structure, similar to structures found in other glyc
osyltransferases, consisting of a short N-terminal cytoplasmic domain,
a signal-membrane anchor domain, a proteolytically sensitive stem reg
ion and a large C-terminal active domain. The identity of this enzyme
was confirmed by construction of a recombinant sialyltransferase in wh
ich the N-terminus part including the cytoplasmic tail, signal anchor
domain and stem region was replaced with an immunoglobulin signal pept
ide sequence. The expression of this recombinant protein in COS-7 cell
s resulted in secretion of a catalytically active and soluble form of
the enzyme into the medium. The expressed enzyme exhibited activity on
ly towards the disaccharide moiety of Gal beta 1,4GlcNAc in glycoprote
ins.