Production and purification of a recombinant human 14 kDa beta-galactoside-binding lectin.
The cDNA for a 14 kDa human beta-galactoside-binding lectin was inserted into a plasmid carrying a taq promoter, and the lectin protein was expressed in E. coli cells. The recombinant lectin was extracted from the cells and purified to apparent homogeneity by a single-step chromatography on an asialofetuin-agarose column. Subunit molecular mass (14 kDa), hemagglutinating activity and antigenicity were indistinguishable from those of the human placental lectin. Though the N-terminal of the placental lectin is blocked with an acetyl group, the recombinant lectin was found to have a free amino group. However, the N-terminal amino acid sequences were identical. The recombinant lectin was considered to have the same three-dimensional structure as the placental lectin.
Hirabayashi
J
J
Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Kanagawa, Jappan.
Ayaki
H
H
Soma
G
G
Kasai
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005697:Galactosides / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D037161:Galectins; D006027:Glycosides / Q000302:isolation & purification*; D006388:Hemagglutinins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D005697:Galactosides; D037161:Galectins; D006027:Glycosides; D006388:Hemagglutinins; D011994:Recombinant Proteins; D004247:DNA