Expression and purification of exendin-4 dimer in Escherichia coli and its interaction with GLP-1 receptor in vitro.
Exendin-4 is a 39 amino acid peptide isolated from the Gila monster salivary gland. It is 53% homologous to GLP-1 and exhibits similar glucoregulatory activities. In this study, exendin-4 dimer (D-Ex4) was constructed, cloned into plasmid pET32a(+) and expressed in E. coli BL21(DE3). The fusion protein with His-tag at the N-terminus was purified with a Ni-NTA-agarose column. After proteolytic cleavage, D-Ex4 peptide with high purity was obtained by HPLC. The results obtained by chemical cross-linking showed that D-Ex4 maintained affinity to GLP-1 receptor.
Yi
Lina
L
State Key Laboratory of Bioreactor Engineering, Institute of Biochemistry, East China University of Science and Technology, Shanghai, 200237, China.
Yin
Xiaopu
X
Wei
Dongzhi
D
Ma
Yushu
Y
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D000077270:Exenatide; D015870:Gene Expression / Q000235:genetics; D000067757:Glucagon-Like Peptide-1 Receptor; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D018027:Receptors, Glucagon / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism*; D014688:Venoms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000067757:Glucagon-Like Peptide-1 Receptor; D010455:Peptides; D018027:Receptors, Glucagon; D011993:Recombinant Fusion Proteins; D014688:Venoms; D000077270:Exenatide