Reduced temperature dependence of collective conformational opening in a hyperthermophile rubredoxin.
Spatially localized differences in the conformational dynamics of the rubredoxins from the hyperthermophile Pyrococcus furiosus (Pf) and the mesophile Clostridium pasteurianum (Cp) are monitored via amide exchange measurements. As shown previously for the hyperthermophile protein, nearly all backbone amides of the Cp rubredoxin exhibit EX(2) hydrogen exchange kinetics with conformational opening rates of >1 s(-)(1). Significantly slower amide exchange is observed for Pf rubredoxin in the region surrounding the metal site and the proximal end of the three-stranded beta-sheet, while for the rest of the structure, the exchange rates at 23 degrees C are similar for both proteins. For the multiple-turn region comprising residues 14-32 in both rubredoxins, the uniformity of both the exchange rate constants and the values of the activation energy at the slowly exchanging sites is consistent with a model of solvent exposure via a subglobal cooperative conformational opening. In contrast to the common expectation of increased rigidity in the hyperthermophile proteins, below room temperature Pf rubredoxin exhibits a larger apparent flexibility in this multiple-turn region. The smaller enthalpy for the conformational opening process of this region in Pf rubredoxin reflects the much weaker temperature dependence of the underlying conformational equilibrium in the hyperthermophile protein.
Hernández
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Wadsworth Center and Department of Biomedical Sciences, University at Albany-State University of New York, Empire State Plaza, Albany, New York 12201-0509, USA.
LeMaster
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DM
D003013:Clostridium / Q000737:chemistry; D003198:Computer Simulation; D004355:Drug Stability; D004563:Electrochemistry; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D019711:Pyrococcus / Q000737:chemistry; D012416:Rubredoxins / Q000737:chemistry*; D013816:Thermodynamics
D012416:Rubredoxins