Poly(L–lysine)–DNA interactions in NaCl solutions: B → C and B → ψ Transitions

✍ Scribed by Manuel Weiskopf; Hsueh Jei Li

• Publisher: Wiley (John Wiley & Sons)
• Year: 1977
• Tongue: English
• Weight: 940 KB
• Volume: 16
• Category: Article
• ISSN: 0006-3525
• DOI: 10.1002/bip.1977.360160315

✦ Synopsis

Abstract

Thermal denaturation and circular dichroism (CD) properties of poly(L‐lysine)–DNA complexes vary greatly when these complexes are prepared differently, that is, whether by NaCl‐gradient dialysis starting from 2.0 M NaCl or by direct mixing at low salt. These differing properties were investigated in more detail by examining complexes, made by direct mixing in the presence of various concentrations of NaCl, both before and after the NaCl was dialyzed out of the complex solution. The precipitation curves of DNA due to polylysine binding indicate that such binding is noncooperative at zero salt; from 0.1 up to 1.0 M NaCl they exhibit varying degrees of cooperatively. Starting from zero salt, as the NaCl concentration used for complex formation is increased, both the CD and the melting properties of the complexes are shifted from those of directly mixed at zero salt to those of reconstitution: in the CD spectra there is a gradual shift from a B → C transition to a B → ψ transition; thermal denaturation results show a gradual increase in the melting temperatures of both free DNA (t~m~) and polylysine‐bound DNA (t′~m~). The progressive shift from B → C to B → ψ suggests a close relationship between these two transitions. Large aggregates of the complexes do not warrant the appearance of ψ‐type CD spectra: ψ‐spectra have been obtained in the supernatants of polylysine–DNA complexes made and measured at 1.0 M NaCl while slightly perturbed CD spectra in B → C transition have been observed in turbid solutions of fully covered complexes made at very low salt. If the complexes are made at intermediate salts and dialyzed to a very low salt, although up to 60% of the DNA is still bound by polylysine, the CD spectra of the complexes are shifted back to the B‐type CD characteristic of pure DNA.