Bacterially expressed rat retinol-binding protein is functional for retinol and transthyretin bindings.
Retinol-binding protein (RBP) was expressed in Escherichia coli using the cDNA for rat RBP, and characterized. The expressed RBP was fused to maltose-binding protein (MBP) at the N-terminal end (MBP-RBP), and MBP was enzymatically removed from the MBP-RBP with proteinase factor Xa. The binding of retinol and transthyretin (TTR) to the recombinant RBP was monitored by means of gel filtration. The recombinant RBP specifically bound to retinol with an affinity similar to that of purified RBP from rat serum. Furthermore, the retinol-bound recombinant RBP formed hetero-complexes with TTR similar to RBP. Thus, the results showed that the recombinant RBP expressed in E. coli is as functional as serum RBP in terms of retinol and TTR bindings.
Yamamoto
Y
Y
Department of Agricultural Chemistry, Faculty of Agriculture, Tokyo University of Agriculture, Japan.
Yoshizawa
T
T
Mano
H
H
Masushige
S
S
Kato
S
S
D018528:ATP-Binding Cassette Transporters; D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D015951:Factor Xa / Q000378:metabolism; D015870:Gene Expression; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011228:Prealbumin / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D014801:Vitamin A / Q000378:metabolism*
D018528:ATP-Binding Cassette Transporters; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011228:Prealbumin; D011993:Recombinant Fusion Proteins; D012177:Retinol-Binding Proteins; C461599:maltose transport system, E coli; D014801:Vitamin A; D015951:Factor Xa