Development of pituitary adenylate cyclase activating polypeptides (PACAPs) specific radioimmunoassay systems and distribution of PACAP-like immunoreactivity in guinea pig tissues.

Biomedical peptides, proteins & nucleic acids : structure, synthesis & biological activity Pub Date : 1994-01-01

E Ando, K Nokihara, S Naruse

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Abstract

Pituitary adenylate cyclase activating polypeptide (PACAP) specific radioimmunoassay systems have been developed and the distribution of PACAPs in guinea pig tissues has been studied. The antibody against PACAP38 was characterized, using synthetic peptide fragments. It was shown to recognize the C-terminal portion with the C-terminal amide group and no cross-reaction was observed with vasoactive intestinal polypeptide (VIP), which has a high homology with PACAP27 and the N-terminal 28 amino acid residues of PACAP38. The antibody against PACAP27 was specific to PACAP27. With the two PACAPs (PACAP38 and PACAP27) specific RIA systems, high concentrations of PACAP38- and PACAP27-like immunoreactivity (LI) were observed in the brain of guinea pigs, especially in the diencephalon (mostly hypothalamus). In all tissues PACAP38-LI was higher than that of PACAP27-LI. Reverse-phase HPLC showed that PACAP38- and PACAP27-LI of tissue extracts were superimposed at the elution position of those of the synthetic peptides, respectively.

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垂体腺苷酸环化酶激活多肽(PACAPs)特异性放射免疫测定系统的建立及pacap样免疫反应性在豚鼠组织中的分布

垂体腺苷酸环化酶激活多肽(PACAP)特异性放射免疫测定系统已经开发，并研究了PACAP在豚鼠组织中的分布。利用合成肽段对该抗体进行了鉴定。与PACAP27和PACAP38的n -末端28个氨基酸残基具有高度同源性的血管活性肠多肽(VIP)未发生交叉反应。抗PACAP27的抗体对PACAP27具有特异性。使用两种PACAP38和PACAP27特异性RIA系统，在豚鼠大脑中观察到高浓度的PACAP38-和PACAP27样免疫反应性(LI)，特别是在间脑(主要是下丘脑)。在所有组织中，PACAP38-LI均高于PACAP27-LI。反相高效液相色谱法显示，组织提取物的PACAP38-和PACAP27-LI分别叠加在合成肽的洗脱位置。

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Biomedical peptides, proteins & nucleic acids : structure, synthesis & biological activity

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