Walker B, Gray J, Burns D M, Wang Q, Adrian T E, Nichols D H, Murphy R F, Nelson J
Division of Biochemistry, Queen's University of Belfast, N. Ireland, UK.
Peptides. 1995;16(2):255-61. doi: 10.1016/0196-9781(94)00174-x.
Two neuromedin C (NC) analogues were constructed by Fmoc synthesis and in situ coupling of 4(5)-carboxyfluorescein or biotin to the N-terminus. Both displayed full agonism in an amylase release assay and cross-reacted fully with a NC-specific antiserum. Biotin NC functioned in a streptavidin-capture ELISA. Carboxyfluorescein NC was used to probe receptor localization in rat stomach. Specific NC binding sites, which did not interact with substance P, angiotensin I, or neurokinin A, were labeled in the antrum. Identity of NC binding sites was confirmed by microautoradiography. The specifically labeled cells were all found in the lamina propria and at least some of cells were identified as eosinophils.
通过Fmoc合成法构建了两种神经介素C(NC)类似物,并将4(5)-羧基荧光素或生物素原位偶联至其N端。二者在淀粉酶释放试验中均表现出完全激动活性,且能与NC特异性抗血清发生完全交叉反应。生物素化NC可用于链霉亲和素捕获ELISA。羧基荧光素标记的NC用于探测大鼠胃中受体的定位。在胃窦中标记出了不与P物质、血管紧张素I或神经激肽A相互作用的特异性NC结合位点。通过显微放射自显影法证实了NC结合位点的特性。特异性标记的细胞均位于固有层,且至少部分细胞被鉴定为嗜酸性粒细胞。