Characterization of rat N-acetylglucosaminyltransferase I expressed in Escherichia coli.
The catalytic domain (sGnT-I) of rat liver N-acetylglucosaminyl-transferase I (GnT-I) was expressed in Escherichia coli. Lysates from pETsGnT-I transformants contained a prominent protein species of 46 kDa with which a significant GnT-I activity was associated. To purify the relevant enzyme, we constructed cDNAs encoding sGnT-ICH and sGnT-INH, which had six additional histidine residues as an affinity tag at the C-terminal and the N-terminal of sGnT-I, respectively, and introduced them into E. coli cells for expression. sGnT-INH was purified and its enzymatic properties were examined.
Nishiu
J
J
Department of Agricultural Chemistry, Kyoto University, Japan.
Kioka
N
N
Fukada
T
T
Sakai
H
H
Komano
T
T
D000818:Animals; D002240:Carbohydrate Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011134:Polysaccharides / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis
D011134:Polysaccharides; D011994:Recombinant Proteins; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I