Covalent affinity labeling of the formyl peptide chemotactic receptor.
Covalent affinity labeling of the formyl peptide chemotactic receptor.
复制标题
甲酰肽趋化受体的共价亲和标记。
DOI：
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发表时间：
1980
影响因子：
4.8
通讯作者：
P. Cuatrecasas
中科院分区：
文献类型：
--
作者：
J. Niedel;J. Davis;P. Cuatrecasas
甲酰基肽趋化受体目前在人中性粒细胞膜上已共价亲和标记使用三种不同的技术。N-甲酰基-Nle-Leu-Phe-Nle-125 I-Tyr-Lys与受体通过辛二酰亚胺二甲酯交联，N-甲酰基-Nle-Leu-Phe-Nle-125 I-Tyr-Lys-Nepropyl-4-叠氮基-2-硝基苯基通过光活化交联，N-甲酰基-Nle-Leu-Phe-Nle-125 I-Tyr-Lys-Nepropyl-bromoacetyl与受体自发反应。用每种方法，特异性标记在十二烷基硫酸钠-聚丙烯酰胺电泳上迁移为宽带的多肽，其表观分子量在55，000和70，000之间。未标记的N-甲酰基-Nle-Leu-Phe-Nle-Tyr-Lys抑制甲酰基肽衍生物共价交联的剂量-反应曲线与抑制相同甲酰基肽衍生物与受体结合的剂量-反应曲线密切相关。非甲酰化类似物Nle-Leu-Phe-Nle-Tyr-Lys不抑制结合或共价交联。
The formyl peptide chemotactic receptor present on human neutrophil membranes has been covalently affinity-labeled using three different techniques. N-Formyl-Nle-Leu-Phe-Nle-125I-Tyr-Lys was cross-linked to the receptor with dimethyl suberimidate, N-formyl-Nle-Leu-Phe-Nle-125I-Tyr-Lys-Nepsilon-4-azido-2-nitrophenyl was cross-linked by photoactivation, and N-formyl-Nle-Leu-Phe-Nle-125I-Tyr-Lys-Nepsilon-bromoacetyl reacted spontaneously with the receptor. With each method, a polypeptide which migrated as a broad band on sodium dodecyl sulfate-polyacrylamide electrophoresis, with an apparent molecular weight between 55,000 and 70,000, was specifically labeled. A dose-response curve for inhibition of covalent cross-linking of the formyl peptide derivatives by unlabeled N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys correlated closely with a dose-response curve for inhibition of binding of the same formyl peptide derivatives to the receptor. The nonformylated analog, Nle-Leu-Phe-Nle-Tyr-Lys, did not inhibit binding or covalent cross-linking.