Expression, purification and binding to the receptor of human insulin-like growth factor II.
Human insulin-like growth factor II (IGF-II) was expressed as a fused protein with 14 additive amino acids in Escherichia coli with a high yield by an expression system using T7 RNA polymerase. Purification of the expressed protein was simply performed using only differential ultrafiltrations, giving a homogeneous preparation upon polyacrylamide gel electrophoresis and high-performance liquid chromatography. The expressed peptide was reacted with a monoclonal antibody raised against native IGF-II on a blotted membrane. Furthermore, the peptide was bound to IGF-II receptor in solubilized rat fetus membrane, though the affinity was slightly inferior to that of native IGF-II. In addition, fusion IGF-II immobilized on a gel matrix was useful for one-step purification of the IGF-II receptor with a high yield from solubilized rat fetus membranes.
Miyagishima
T
T
Biochemistry Laboratory Cancer Institute, Hokkaido University School of Medicine, Sapporo, Japan.
Gasa
S
S
Honke
K
K
Sakai
M
M
Nishi
S
S
Yamamoto
M
M
Nishikawa
K
K
Miyazaki
T
T
Makita
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004926:Escherichia coli / Q000378:metabolism; D005321:Extraembryonic Membranes / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D051381:Rats; D017527:Receptor, IGF Type 2 / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification
D017527:Receptor, IGF Type 2; D011993:Recombinant Fusion Proteins; D007335:Insulin-Like Growth Factor II