Hyper-mobility of water around actin filaments revealed using pulse-field gradient spin-echo 1H NMR and fluorescence spectroscopy

被引:9
作者
Wazawa, Tetsuichi [1 ,2 ]
Sagawa, Takashi [1 ]
Ogawa, Tsubasa [1 ]
Morimoto, Nobuyuki [1 ]
Kodama, Takao [3 ]
Suzuki, Makoto [1 ,2 ]
机构
[1] Tohoku Univ, Grad Sch, Dept Mat Proc, Aoba Ku, Sendai, Miyagi 9808579, Japan
[2] JST, CREST, Kawaguchi, Saitama 3320012, Japan
[3] Osaka Univ, Immunol Frontier Res Ctr, Suita, Osaka 5650871, Japan
关键词
Self-diffusion; Fluorescence anisotropy; Rotational correlation time; Local viscosity; Structure-breaker; Hyper-mobile water; Endothermic process; Actin polymerization; F-ACTIN; SELF-DIFFUSION; POLYMERIZATION; POLYMERS; MYOSIN;
D O I
10.1016/j.bbrc.2010.12.096
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
This paper reports that water molecules around F-actin, a polymerized form of actin, are more mobile than those around G-actin or in bulk water. A measurement using pulse-field gradient spin-echo H-1 NMR showed that the self-diffusion coefficient of water in aqueous F-actin solution increased with actin concentration by similar to 5%, whereas that in G-actin solution was close to that of pure water. This indicates that an F-actin/water interaction is responsible for the high self-diffusion of water. The local viscosity around actin was also investigated by fluorescence measurements of Cy3, a fluorescent dye, conjugated to Cys 374 of actin. The steady-state fluorescence anisotropy of Cy3 attached to F-actin was 0.270, which was lower than that for G-actin, 0.334. Taking into account the fluorescence lifetimes of the Cy3 bound to actin, their rotational correlation times were estimated to be 3.8 and 9.1 ns for F- and G-actin, respectively. This indicates that Cy3 bound to F-actin rotates more freely than that bound to G-actin, and therefore the local water viscosity is lower around F-actin than around G-actin. (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码:985 / 990
页数:6
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