Direct Evidence for the Effect of Glycerol on Protein Hydration and Thermal Structural Transition

被引:45
作者
Hirai, Mitsuhiro [1 ]
Ajito, Satoshi [1 ]
Sugiyama, Masaaki [2 ]
Iwase, Hiroki [3 ]
Takata, Shin-ichi [4 ]
Shimizu, Nobutaka [5 ]
Igarashi, Noriyuki [5 ]
Martel, Anne [6 ]
Porcar, Lionel [6 ]
机构
[1] Gunma Univ, Grad Sch Sci & Technol, Maebashi, Gunma, Japan
[2] Kyoto Univ, Inst Res Reactor, Osaka, Japan
[3] Comprehens Res Org Sci & Soc, Tokai, Ibaraki, Japan
[4] Japan Atom Energy Agcy, J PARC Ctr, Tokai, Ibaraki, Japan
[5] High Energy Accelerator Res Org, Inst Mat Struct Sci, Tsukuba, Ibaraki, Japan
[6] Inst Laue Langevin, Grenoble, France
关键词
PREFERENTIAL INTERACTION COEFFICIENTS; X-RAY-SCATTERING; NEUTRON-SCATTERING; AMYLOID FIBRILS; DYNAMICS; MYOGLOBIN; WATER; STABILITY; APOMYOGLOBIN; SOLVATION;
D O I
10.1016/j.bpj.2018.06.005
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The mechanisms of protein stabilization by uncharged solutes, such as polyols and sugars, have been intensively studied with respect to the chemical thermodynamics of molecular crowding. In particular, many experimental and theoretical studies have been conducted to explain the mechanism of the protective action on protein structures by glycerol through the relationship between hydration and glycerol solvation on protein surfaces. We used wide-angle x-ray scattering (WAXS), small-angle neutron scattering, and theoretical scattering function simulation to quantitatively characterize the hydration and/or solvation shell of myoglobin in aqueous solutions of up to 75% v/v glycerol. At glycerol concentrations below similar to 40% v/v, the preservation of the hydration shell was dominant, which was reasonably explained by the preferential exclusion of glycerol from the protein surface (preferential hydration). In contrast, at concentrations above 50% v/v, the partial penetration or replacement of glycerol into or with hydration-shell water (neutral solvation by glycerol) was gradually promoted. WAXS results quantitatively demonstrated the neutral solvation, in which the replacement of hydrated water by glycerol was proportional to the volume fraction of glycerol in the solvent multiplied by an exchange rate (beta <= 1). These phenomena were confirmed by small-angle neutron scattering measurements. The observed WAXS data covered the entire hierarchical structure of myoglobin, ranging from tertiary to secondary structures. We separately analyzed the effect of glycerol on the thermal stability of myoglobin at each hierarchical structural level. The thermal transition midpoint temperature at each hierarchical structural level was raised depending on the glycerol concentration, with enhanced transition cooperativeness between different hierarchical structural levels. The onset temperature of the helix-to-cross beta-sheet transition (the initial process of amyloid formation) was evidently elevated. However, oligomerization connected to fibril formation was suppressed, even at a low glycerol concentration.
引用
收藏
页码:313 / 327
页数:15
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