Relationship between protein stabilization and protein rigidification induced by mannosylglycerate.
Understanding protein stabilization by small organic compounds is a topic of great practical importance. The effect of mannosylglycerate, a charged compatible solute typical of thermophilic microorganisms, on a variant of staphylococcal nuclease was investigated using several NMR spectroscopy methods. No structural changes were apparent from the chemical shifts of amide protons. Measurements of (15)N relaxation and model-free analysis, water-amide saturation transfer (phase-modulated CLEAN chemical exchange), and hydrogen/deuterium exchange rates provided a detailed picture of the effects of mannosylglycerate on the backbone dynamics and time-averaged structure of this protein. The widest movements of the protein backbone were significantly constrained in the presence of mannosylglycerate, as indicated by the average 5-fold decrease of the hydrogen/deuterium exchange rates, but the effect on the millisecond timescale was small. At high frequencies, internal motions of staphylococcal nuclease were progressively restricted with increasing concentrations of mannosylglycerate or reduced temperature, while the opposite effect was observed with urea (a destabilizing solute). The order parameters showed a strong correlation with the changes in the T(m) values induced by different solutes, determined by differential scanning calorimetry. These data show that mannosylglycerate caused a generalised reduction of backbone motions and demonstrate a correlation between protein stabilization and protein rigidification.
Pais
Tiago M
TM
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Rua da Quinta Grande 6, Apartado 127, 2780-156 Oeiras, Portugal.
Lamosa
Pedro
P
Garcia-Moreno
Bertrand
B
Turner
David L
DL
Santos
Helena
H
D000577:Amides / Q000737:chemistry; D002152:Calorimetry, Differential Scanning; D003903:Deuterium / Q000737:chemistry; D005988:Glyceric Acids / Q000737:chemistry*; D006859:Hydrogen / Q000737:chemistry; D008358:Mannose / Q000031:analogs & derivatives* / Q000737:chemistry; D008836:Micrococcal Nuclease / Q000737:chemistry; D017510:Protein Folding; D055550:Protein Stability; D014867:Water / Q000737:chemistry
D000577:Amides; D005988:Glyceric Acids; C402688:mannosylglycerate; D014867:Water; D006859:Hydrogen; D003903:Deuterium; D008836:Micrococcal Nuclease; D008358:Mannose