Yeung C W
Int J Pept Protein Res. 1985 Jul;26(1):63-9.
A photoreactive [D-Ala2, p-N3-Phe4-Met5]enkephalin was synthesized by classical solution peptide synthetic methods. The hydroxysuccinimide ester was used in all the coupling steps in the presence of a weak base, triethylamine. The deprotected enkephalin analogue was purified on high performance liquid chromatography using a Waters, C18 muBondapak reverse phase column and its purity was assessed by thin-layer chromatography. The composition of the analogue was determined and confirmed by elemental analysis and amino acid analysis. Its photoreactivity was demonstrated by the time dependent ultraviolet spectral changes on exposure to light. Competition receptor binding showed that [D-Ala2, p-N3-Phe4-Met5]enkephalin was 4-fold more potent than [D-Ala2, Met5]-enkephalin in competing for binding to the enkephalin binding site. The data presented suggest that this photoreactive enkephalin analogue may be suitable for use in the in situ photoaffinity labeling of the enkephalin receptor.
一种光反应性的[D - Ala2, p - N3 - Phe4 - Met5]脑啡肽通过经典的溶液肽合成方法合成。在弱碱三乙胺存在的情况下，羟基琥珀酰亚胺酯用于所有偶联步骤。脱保护的脑啡肽类似物使用沃特世C18 μBondapak反相柱通过高效液相色谱法进行纯化，其纯度通过薄层色谱法评估。该类似物的组成通过元素分析和氨基酸分析确定并确认。其光反应性通过暴露于光时随时间变化的紫外光谱变化得以证明。竞争受体结合实验表明，[D - Ala2, p - N3 - Phe4 - Met5]脑啡肽在竞争结合脑啡肽结合位点方面比[D - Ala2, Met5] - 脑啡肽的效力高4倍。所呈现的数据表明，这种光反应性脑啡肽类似物可能适用于脑啡肽受体的原位光亲和标记。
