Intein-mediated rapid purification of recombinant maxadilan and M65 and their acute effects on plasma glucose  

作　　者：Rongjie Yu Tianhong Yi Ling Zhang An Hong Yun Dai Tianhong Zhou 

机构地区：[1]Bio-engineering Institute of Jinan University, Jinan University, Guangzhou 510632, China [2]Bio-engineering Institute of Jinan University, Life Science Department, Jinan University, Guangzhou 510632, China

出　　处：《Acta Biochimica et Biophysica Sinica》2008年第12期1015-1022,共8页生物化学与生物物理学报（英文版）

基　　金：This work was supported by grants from the Guangdong Provincial Nature Science Foundation of China （No. 06300579）, the Science Technology Research Key Project of the Educational Department of China （No. 207141） and the National Technology Development Plan （No. 2006AA02Z125）

摘　　要：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 chitinbound intein tag by β-mercaptoethanol. Intraperitoneal injection of the recombinant maxadilan caused an acute elevation of plasma glucose, imitating pituitary adenylate cyclaseactivating 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.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 chitinbound intein tag by β-mercaptoethanol. Intraperitoneal injection of the recombinant maxadilan caused an acute elevation of plasma glucose, imitating pituitary adenylate cyclaseactivating 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.

关 键 词：PURIFICATION MAXADILAN PAC1 INTEIN plasmaglucose 

分 类 号：R32[医药卫生—人体解剖和组织胚胎学]