Comparison of the internal dynamics of globular proteins in the microcrystalline and rehydrated lyophilized states.
Natural abundance solid-state 13C-NMR spin-lattice relaxation experiments in the laboratory (T1) and off-resonance rotating (T(1rho)) frames were applied for qualitative comparison of the internal molecular dynamics of barstar, hen egg white lysozyme and bacteriophage T4 lysozyme in both the microcrystalline and the rehydrated (water content is 50% of the protein mass) lyophilized states. The microcrystalline state of proteins provides a better spectral resolution; however, less is known about the local structure and dynamics in the different states. We found by visual comparison of both T1 and T(1rho) relaxation decays of various resonance bands of the CPMAS spectra that within the ns-mus range of correlation times there is no appreciable difference in the internal dynamics between rehydrated lyophilized and crystalline states for all three proteins tested. This suggests that the internal conformational dynamics depends weakly if at all on inter-protein interactions in the solid state. Hence, physical properties of globular proteins in a fully hydrated solid state seem to be similar to those in solution. This result at least partly removes concerns about biological relevance of studies of globular proteins in the solid state.
Krushelnitsky
Alexey
A
Kazan Institute of Biochemistry and Biophysics, Kazan, Russia. krushelnitsky@mail.knc.ru
Gogolev
Yuri
Y
Golbik
Ralph
R
Dahlquist
Frederick
F
Reichert
Detlef
D
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D003461:Crystallography; D005612:Freeze Drying; D009113:Muramidase / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D014764:Viral Proteins / Q000737:chemistry*
D001426:Bacterial Proteins; D014764:Viral Proteins; C068293:gene 5 protein, Enterobacteria phage T4; C057438:barstar protein, Bacillus amyloliquefaciens; C421864:hen egg lysozyme; D009113:Muramidase