Functional characterization of structural alterations in the sequence of the vasodilatory peptide maxadilan yields a pituitary adenylate cyclase-activating peptide type 1 receptor-specific antagonist
Functional characterization of structural alterations in the sequence of the vasodilatory peptide maxadilan yields a pituitary adenylate cyclase-activating peptide type 1 receptor-specific antagonist
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DOI：
10.1074/jbc.274.33.23103
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发表时间：
1999-08-13
影响因子：
4.8
通讯作者：
Tajima, M
中科院分区：
文献类型：
--
作者：
Moro, O;Wakita, K;Tajima, M
Maxadilan是一种从沙蝇中提取的血管扩张肽，是垂体腺苷酸环化酶激活肽(PACAP)1型受体的激动剂。令人惊讶的是，Maxadilan与PACAP没有显著的序列同源性，为了研究Maxadilan的结构和活性之间的关系，在多肽中进行了几个氨基酸的替换和缺失。体外检测这些多肽与表达PACAP 1型受体的兔脑组织的粗膜结合；在表达这些受体的PC12细胞中检测cAMP的诱导；体内检测这些多肽诱导兔皮肤红斑的能力。取代1和5位上的单个半胱氨酸或删除该环结构对活性几乎没有影响。第14位或51位半胱氨酸的取代消除了活性。缺失第24位和42位之间的19个氨基酸，得到一个具有结合活性但没有功能活性的多肽。该缺失突变体与转染PACAP 1型受体的COS细胞的相互作用表明，该多肽是PACAP 1型受体的特异性拮抗剂。
Maxadilan is a vasodilatory peptide derived from sand flies that is an agonist at the pituitary adenylate cyclase-activating peptide (PACAP) type 1 receptor. Surprisingly, maxadilan does not share significant sequence homology with PACAP, To examine the relationship between structure and activity of maxadilan, several amino acid substitutions and deletions were made in the peptide. These peptides were examined in vitro for binding to crude membranes derived from rabbit brain, a tissue that expresses PACAP type 1 receptors; and induction of cAMP was determined in PC12 cells, a line that expresses these receptors, The peptides were examined in vivo for their ability to induce erythema in rabbit skin. Substitution of the individual cysteines at positions 1 and 5 or deletion of this ring structure had little effect on activity. Substitution of either cysteine at position 14 or 51 eliminated activity. Deletion of the 19 amino acids between positions 24 and 42 resulted in a peptide with binding, but no functional activity. The capacity of this deletion mutant to interact with COS cells transfected with the PACAP type 1 receptor revealed that this peptide was a specific antagonist to the PACAP type 1 receptor.