Application of a lanthanide tag for evaluation of conformational states of a multidomain protein

Saio, Tomohide*; Nakagawa, Hiroshi   ; Hiramatsu, Soya*; Asada, Mizue*; Kawamukai, Honoka*; Nakamura, Toshikazu*; Ishimori, Koichiro*

Despite their importance in function, the conformational states and changes of proteins are often poorly understood mainly because of the lack of an efficient tool. MurD, a 47-kDa three-domain protein enzyme responsible for peptidoglycan biosynthesis, is one of those proteins whose conformational states and changes during its catalytic cycle are not well understood. We exploited multiple biophysical methods including nuclear magnetic resonance, electron paramagnetic resonance, small-angle scattering, and molecular dynamics simulation to demonstrate evaluation of the conformational states and distribution of MurD. The integrated use of these methods can be an efficient strategy to evaluate the conformational states and distribution of proteins in solution.