Fréchet D, Guitton J D, Herman F, Faucher D, Helynck G, Monegier du Sorbier B, Ridoux J P, James-Surcouf E, Vuilhorgne M
Rhône-Poulenc Rorer S.A., Vitry-Sur-Seine, France.
Biochemistry. 1994 Jan 11;33(1):42-50. doi: 10.1021/bi00167a006.
The structure of RP 71955, a new tricyclic 21 amino acid peptide active against human immunodeficiency virus 1, was determined. Its amino acid composition was inferred from the results of fast atom bombardment mass spectrometry, nuclear magnetic resonance, Raman spectroscopy, and amino acid analysis. Its sequence could not be determined classically, using Edman degradation, given the lack of a free terminal NH2. It was deduced from the interpretation of interresidue nuclear Overhauser effects and confirmed by the sequencing of peptides obtained by limited chemical hydrolysis. It was found to be CLGIGSCNDFAGCGYAVVCFW. An internal amide bond between the NH2 of C1 and the gamma-COOH of D9 was observed, as well as two disulfide bridges, one between C1 and C13 and one between C7 and C19. The three-dimensional structure of RP 71955 was determined from nuclear magnetic resonance derived constraints using distance geometry, restrained molecular dynamics, nuclear Overhauser effect back calculation, and an iterative refinement using a full relaxation matrix approach. Analogies between the structure of RP 71955 and some functional domains of gp41, the transmembrane protein of human immunodeficiency virus 1, suggest hypotheses concerning the mode of action of RP 71955.
确定了新型三环21氨基酸肽RP 71955的结构，该肽对人免疫缺陷病毒1具有活性。其氨基酸组成是根据快原子轰击质谱、核磁共振、拉曼光谱和氨基酸分析结果推断出来的。由于缺乏游离的末端NH2，无法使用埃德曼降解法经典地确定其序列。它是通过对残基间核Overhauser效应的解释推导出来的，并通过有限化学水解得到的肽段测序得到证实。结果发现其序列为CLGIGSCNDFAGCGYAVVCFW。观察到C1的NH2与D9的γ-COOH之间存在一个内部酰胺键，以及两个二硫键，一个在C1和C13之间，另一个在C7和C19之间。利用距离几何、受限分子动力学、核Overhauser效应反算以及使用全松弛矩阵方法的迭代优化，从核磁共振衍生的约束条件确定了RP 71955的三维结构。RP 71955的结构与人类免疫缺陷病毒1的跨膜蛋白gp41的一些功能域之间的相似性，提出了关于RP 71955作用模式的假设。
