Hyper-Mobile Water around Ions, Charged Polymers, and Proteins Observed with High Resolution Microwave Dielectric Spectroscopy
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1 Netsu Sokutei Hyper-Mobile Water around Ions, Charged Polymers, and Proteins Observed with High Resolution Microwave Dielectric Spectroscopy Makoto Suzuki and Takashi Miyazaki (Received October 1, 2007; Accepted October 17, 2007) A precision microwave dielectric spectroscopic technique has been developed. This technique enables us to analyze hydration properties of ions, polymers, and proteins. φ -scan method gives an image of the information of spatial distribution of dielectric property of water surrounding protein molecules. Fixed φ analysis gives a direct comparison of hydration properties of different solutes. With this technique we analyzed the hydration states of alkali-halides aqueous solutions, charged polymers and proteins such as F- actin solutions. Most of solutes called water structure breakers form hyper-mobile water (HMW) layer around the solutes having higher dielectric relaxation frequency than bulk water. Thus, we may obtain further information of collective properties of water surrounding ions, polymers and biomolecules. Keywords: Microwave dielectric spectroscopy; protein hydration; actin; hyper-mobile water; φ -scan method; dielectric property of water 1950 Samoirov 1) Frank Wen 2) water structure breakers chaotropes negative hydration Hofmeister 3) 4) 5) 2007 The Japan Society of Calorimetry and Thermal Analysis. 244
2 Hyper-mobile water found around charged solutes in water GHz hyper-mobile water, HMW water structure breaker HMW HMW GHz 6) γ GHz β 10 7 s 10 6 Hz 0.1 GHz δ 7) 4 8 GHz δ 20 mg ml 1 6-9) Asami 8) Fig.1 8) % Fig.1 Complex dielectric constant of a mixture of ellipsoids and solvent. 8) Fig.1 first shell φ Fig.1 second shell φ bovine serum albumin (BSA), horse radish peroxidase (HRP), myoglobin (Mb), lysozyme (Lys), hemoglobin (Hb) Fig.2 ε q" 6 7 GHz 10) ε q" 6 GHz 1 GHz a/f a f 1 GHz ε q" 5 GHz 1 GHz 2.5 f c1, δ 1, φ N b N b 55.6 φ 1/c M c M φ 1 3D 245
3 0.14 nm ASA 1 N b3d-cal N b Lys HRP α GHz 11,12) 6 7 GHz 1 Fig.2 φ ml g φ/c ml mg HRP Fig.2(a) Fig.2(b) 10 GHz 1 Lys Hb 10 GHz BSA φ f c, δ, N b 2 φ f c δ f c δ ) Fig.2 (a) real part (φ /c ml mg 1 ) (b) imaginary part Partial dielectric spectra of hydrated globular protein in solution. φ f c δ φ f c, δ 2 φ φ B φ B φ φ φ φ GHz φ 246
4 Hyper-mobile water found around charged solutes in water HMW φ 1970 Giese 13) GHz CsF τ r KCl, KBr, KI, RbCl, CsCl water structure breakers τ r 1/2π f c, f c : F Li, Na 1 26 GHz NaF NaI 0.1 M, 10 Fig.3,4 2 GHz ε q" σ /2πf σ : 1 GHz a/f a 1 GHz a NaF φ ε q' φ 0.1 M φ Na F 1 40 φ δ i/[1 i(f/f ci)] GHz 14.5 GHz 12.6 GHz Fig.3 φ -scan spectra of 0.1 M NaF aqueous solution at 10. Fig.4 φ -scan spectra of 0.1 M NaI aqueous solution at 10. f c HMW 5 NaI 0.1 M φ Na I GHz HMW Na 1 26 GHz NaF HMW Giese NaI HMW HMW1.3 HMW63 247
5 Electric field strength / MV m 1 polymer surfacr (a) Distance / nm Fig.5 Rough estimation of electric field strength around polyionenes. (b) - [(- CH 2 -) x -N (Br )(CH 3) 2] n - - CH 2 - x I HMW MV m 1 HMW x 2, 3, 4, 6 0.1M Fig.5 x 2, 3HMW x 6 x mg ml 1 φ Fig.6 x 2 x Huxley 14) S Huxley Simmons 15) ) 1993 S1 3 17) X 18) Fig.6 φ -scan spectra of polyionene aqueous solution at 20. (a) x 2, (b) x 6, c 20 mg ml ) ATP Kodama 20) S1 ATP Gopal Burke 21) S1 ATPS1 Katoh Morita 22) S1 ATP Fig.7 248
6 Hyper-mobile water found around charged solutes in water Dielectric constant of hydrated protein in solution Fig.7 Partial dielectric spectra of F-actin, myosin S1 and Acto-S1. φ/c ml mg 1, φ : c : mg ml mg ml 1, 2 mm HEPES (ph 7.2), 0.2 mm ATP, 0.1 mm CaCl 2, 50 mm KCl, 2 mm MgCl 2 Fig.7 6 GHz S1 6 GHz 20 GHz 7 GHz S1 20 GHz 7 GHz HMW Fig GHz 1200 δ GHz HMW ) δ 2 75 δ 2 60 δ 2 60 F-actin HMW 30 % 1800 F-actin GHz HMW Thomasson 23) MV m 1 S1 Fig.7 S1 10 GHz φ φ/c ml mg 1 S1 1 7 GHz 17 GHz 10 GHz S GHz HMW F-actin S1 Fig.7 6 GHz F-actin 7 GHz 20 GHz S1, F- S ml mg 1 HMW S1ASA ) ) HMW 1950 Samoilov Frank Wen MV m 1 17 GHz I, Br HMW NMR S1 ATP S1 S1 S1 5,23) HMW 249
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