NMR Studies on YSPTSPSY

Abstract

NMR (600 MHz) studies on YSPTSPSY (1), the heptad repeat unit of RNA polymerase II with an extra tyrosine added to the N-terminus, show that the peptide is partially structured in aqueous solution. Peptide I contains two overlapping SPXX sequences and has been reported to bind to DNA by bisintercalation (Suzuki, M. Nature 1990,344, 562-565). In 90% H2O solution at pH 3.2, the major species, which is present in >90%, contains Pro3 and Pro6 in Idle trans conformation. At low temperature (4 °C), NOE connectivities are consistent with the presence of β-turn structures in equilibrium with unfolded forms of the peptide. A strong dnn connectivity between Thr4/Ser5, a dan connectivity between Pro3/Thr4, and a medium-range NOE between Pro3 a and Ser5 NH indicate the presence of a β-turn formed by (i) Ser2-Pro3-Thr4-Ser5. A strong dNN connectivity between Ser7/Tyr8 and weaker dNδ NOE connectivities between Ser2/Pro3 δ and Ser7/Pro6 δ were also detected. The solution conformation of the peptide appears to have a crucial role in determining the interaction of the peptide with DNA, given that only bisintercalation has been reported in DNA-binding studies on 1 (Suzuki, M. Nature 1990,344,562-565). On the basis of these results, the peptide unit -SPTSPS- (3) has considerable potential as a structured linker in the preparation of DNA bisintercalators of the general structure Xaa-SPTSPS-Zaa (2) with improved selectivity properties. Show more