Open AccessIntein‐mediated rapid purification of recombinant maxadilan and M65 and their acute effects on plasma glucose
Author(s) -
Yu Rongjie,
Yi Tianhong,
Zhang Ling,
Hong An,
Dai Yun,
Zhou Tianhong
Publication year - 2008
Publication title -
acta biochimica et biophysica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.771
H-Index - 57
eISSN - 1745-7270
pISSN - 1672-9145
DOI - 10.1111/j.1745-7270.2008.00485.x
Subject(s) - recombinant dna , intein , agonist , escherichia coli , chemistry , microbiology and biotechnology , adenylate kinase , receptor , medicine , endocrinology , biology , biochemistry , gene , rna , rna splicing
Maxadilan is a potent vasodilatory peptide present in the salivary glands of the sand fly. Maxadilan and M65, a deletion variation of maxadilan, are agonist‐ and antagonist‐specific for the PAC1 receptor. In order to obtain the recombinant maxadilan and M65 efficiently by intein‐mediated single column purification, the genes encoding maxadilan and M65 were designed, synthesized and cloned into Escherichia coli expression vector pKYB. The recombinant maxadilan and M65 with homogeneity over 95% were released from the chitin‐bound intein tag by β‐mercaptoethanol. Intraperitoneal injection of the recombinant maxadilan caused an acute elevation of plasma glucose, imitating pituitary adenylate cyclase‐activating polypeptide (PACAP) 27, in NIH mice, while the VPAC1‐agonist and VPAC2‐agonist had no significant effects on the levels of plasma glucose. M65 alone had no effect on the plasma glucose, but blocked the glucose excursion caused by maxadilan by 12.7% and blocked the glucose excursion caused by the PACAP 27 by 11.6%. The acute effects of the recombinant maxadilan and M65 on the plasma glucose indicated that they had the characteristics as the agonist and antagonist for PAC1.