Geierstanger B H, Volkman B F, Kremer W, Wemmer D E
Graduate Group in Biophysics, University of California, Berkeley 94720.
Biochemistry. 1994 May 3;33(17):5347-55. doi: 10.1021/bi00183a043.
Short peptides derived from chromosomal proteins have previously been proposed to bind specifically to the minor groove of A,T-rich DNA [for a review, see M. E. A. Churchill and A. A. Travers (1991) Trends Biochem. Sci. 16, 92-97]. Using NMR spectroscopy, we investigated the DNA binding of SPRKSPRK, which is one such A,T-specific motif. Under the conditions studied SPRKSPRK interacts only nonspecifically with d(CGCAAAAAAGGC).d(GCCTTTTTTGCG). The peptides TPKRPRGRPKK, PRGRPKK, and PRGRP derived from the non-histone chromosomal protein HMG-I/Y, however, bind specifically to the central A,T sites of d(CGCAAATTTGCG)2 and d(CGCGAATTCGCG)2. 2D NOE measurements show that the RGR segment of each peptide is in contact with the minor groove. The arginine side chains and the peptide backbone are buried deep in the minor groove, in a fashion generally similar to the antibiotic netropsin. Under the same conditions the peptide PKGKP does not interact with the same oligonucleotide duplexes, indicating that the arginine guanidinium groups are major determinants of the A,T specificity.
先前有人提出,源自染色体蛋白的短肽可特异性结合富含A、T的DNA小沟[综述见M. E. A. Churchill和A. A. Travers(1991年)《生物化学趋势》16卷,92 - 97页]。我们利用核磁共振光谱研究了SPRKSPRK的DNA结合情况,它就是这样一个A、T特异性基序。在所研究的条件下,SPRKSPRK仅与d(CGCAAAAAAGGC).d(GCCTTTTTTGCG)发生非特异性相互作用。然而,源自非组蛋白染色体蛋白HMG - I/Y的肽TPKRPRGRPKK、PRGRPKK和PRGRP特异性结合于d(CGCAAATTTGCG)2和d(CGCGAATTCGCG)2的中央A、T位点。二维核欧沃豪斯效应测量表明,每个肽的RGR片段与小沟接触。精氨酸侧链和肽主链深埋于小沟中,其方式总体上类似于抗生素纺锤菌素。在相同条件下,肽PKGKP不与相同的寡核苷酸双链体相互作用,这表明精氨酸胍基是A、T特异性的主要决定因素。