Silvina Cerveny
Material Physics Center, CSIC/UPVSan Sebastian, Spain
Dynamics of water -Case study: molecular relaxations in aqueous solutions of
synthetic and biological materials
Tutorial – BDS 2018
Outline
‐ Where is water? Some fields of research - Confinements
‐ General Phenomenology (glasses, Tg, relaxations) Properties of bulk water / Phase diagram (simulations vs experiments)
-Broadband Dielectric spectroscopy and the study of water molecules
-The dynamics of water at low temperatures in:
- Solution of polymers, glasses and other materials
- Short introduction to Proteins and other biomolecules
- Summary
Where is found water? Where is found water?
Geological waterGeological water
Biological waterBiological water
Water in soft matterWater in soft matter
Where is found water? – Geological waterWhere is found water? – Geological water
Rivers, lakes, reservoirs… BULK LIQUID WATERBULK LIQUID WATER
5 - 10 nm5 - 10 nm5 - 10 nm5 - 10 nm5 - 10 nm
CONFINED WATERCONFINED WATER
Porosity
Some topics of study:
- Water in relation to the structure of rocks as clays, rocks, tobermorite, kaolinite, volcanic ash...-Water motions no crystallization (no man´s land: 150 – 230 K)-How the properties of water change at the nanoscale?
Some topics of study:
- Water in relation to the structure of rocks as clays, rocks, tobermorite, kaolinite, volcanic ash...-Water motions no crystallization (no man´s land: 150 – 230 K)-How the properties of water change at the nanoscale?
Where is found water? – Biological waterWhere is found water? – Biological water
Some topics of study : -The motions of the protein, the hydration shell and the bulk solvent ―> functionality-Analogies confined water in rocks and proteins
Some topics of study : -The motions of the protein, the hydration shell and the bulk solvent ―> functionality-Analogies confined water in rocks and proteins
Most of this water is never more than about 1 nm from other molecules
Most of this water is never more than about 1 nm from other molecules
Model of a bacterial cytoplasm J. Phys. Chem. B 2017, 121
The motions of the protein, the hydration shell and the bulk solvent are all necessary for functionalityThe motions of the protein, the hydration shell and the bulk solvent are all necessary for functionality
Proteins: molecules which perform biological functions
Hydration is necessary for functionalityat least… 0.2 (g of water)/(g of protein) Hydration is necessary for functionalityat least… 0.2 (g of water)/(g of protein)
Bulk solvent
Hydration shellProtein
Where is found water? – Water in all hydrophilic materials, nanocomposites.......Where is found water? – Water in all hydrophilic materials, nanocomposites.......
Hydrophilic polymers contain polar or charged functional groups, rendering them soluble in water.
Some topics of study: -How to dry a polymer?-Plasticization-Compatibility with fillers-……
Some topics of study: -How to dry a polymer?-Plasticization-Compatibility with fillers-……
Some hydrophilic groups:
1. Carboxylic groups: -(COOH)2. Amide: -(C=O)-N-3. Hydroxyl groups: OH-4. .....
Where is found water? Where is found water?
Geological waterGeological water- Confined water (Weathering)- Relationship with structure- Solvent/Buffer
- Confined water (Weathering)- Relationship with structure- Solvent/Buffer
Biological waterBiological water- Bulk water, hydration water,confined water relationship with functionality- Confined water
- Bulk water, hydration water,confined water relationship with functionality- Confined water
Water in soft matterWater in soft matter
- Bulk water, hydration water,confined water- Relationship with several industrial applications
- Bulk water, hydration water,confined water- Relationship with several industrial applications
In many situations water is not in its bulk form, but instead it is either in contact with surfaces or confined within small cavities
Outline
‐ Where is water? Several fields of research. Confinements
‐ General Phenomenology (glasses, Tg, relaxations) Properties of bulk water / Phase diagram (simulations vs experiments)
-Broadband Dielectric spectroscopy and the study of water molecules
-The dynamics of water at low temperatures in:
- Solution of polymers, glasses and other materials
- Short introduction to Proteins and other biomolecules
- Summary
cool down below Tm
crystallizescrystallizes
Fixed positions, minimal potentialEquilibrium state
Does not crystallizesDoes not crystallizes
Supercooled liquid
msNPoise /1010 1213 Liquid is termed a GLASS
Randomly moving
Liquid
Viscosity = (T)Viscosity = (T)
o
oo TT
TDexp Vogel-Fulcher-Tamman (VFT)
1/T
Log()
GlassesGlasses
Glasses – Glass transition temperature - Tg Glasses – Glass transition temperature - Tg
glass
Supe
rcoo
led
liquid
liquid
crystalTM
Tg
T
V
Tg/T
Log()
stron
g
frag
ile
1
o
oo TT
TDexp
T
cp
Tg
273
Supercooled Glassy
T [K]100150235
Crystallization
Bulk water – No man´s land.... Bulk water – No man´s land....
Glassy water (HGW)
106/107 K/sec
Liquid water
Low Density Amorphous ice (LDA)High Density Amorphous ice (HDA)
Vapor/crystalline
(Hallbrucker et all, 1989)
LDA: Sceats and Rice, 1982HDA: Mishima et al, 1984
106 K/sec LDA HDA
No man´s landthe region lying between the crystallization temperatures encountered when heating the glass and cooling the liquid.
??? ???Tg = 136 K?Tg = 200-160 K?
Anomalies of water/ Experiments....Anomalies of water/ Experiments....
Due to crystallization…No entering in the no man´s landDue to crystallization…No entering in the no man´s land
- To mix water with hydrophilic solutes (SOFT CONFINEMENT)
L
L
pores: 2-D confinementlayers: 1-D confinement
- Well-defined confinement systems (HARD CONFINEMENT)
Mineral Clays, Graphite Oxide, Cements, Molecular sieves, MCM 41, Silica hydrogels, Zeolites………
- Polymers - Glass formers materials
Outline
‐ Where is water? Several fields of research. Confinements
‐ General Phenomenology (glasses, Tg, relaxations) Properties of bulk water / Phase diagram (simulations vs experiments)
-Broadband Dielectric spectroscopy and the study of water molecules
-The dynamics of water at low temperatures in:
- Solution of polymers, glasses and other materials
- Short introduction to Proteins and other biomolecules
- Summary
The water moleculeThe water molecule
Water molecule
Hydrogen bond between water molecules BULK Liquid water
Broadband dielectric spectroscopyBroadband dielectric spectroscopy
1) Materials with a permanent dipolar moment
2) External static electric field (1 Volt)
Orientation Polarization
Dielectric spectroscopy can provide information about the segmental mobility of polar moleculesDielectric spectroscopy can provide information about the segmental mobility of polar molecules
3) External electric field is frequency dependent (10-3 to 1011 Hz):
“the dipole relaxation arising from the reorientational motions of molecular dipoles”
Carbon dioxideWater molecule Peptide
Polymers, proteins,….
EEP
o
o
)1(
χ = electric susceptibility = (1+ )
Broadband dielectric spectroscopyBroadband dielectric spectroscopy
100 101 102 103 104 105 106 107 108 109 1010
T1< T2
-relaxation
Hz
-relaxationT1< T2
Broadband dielectric spectroscopyBroadband dielectric spectroscopy Information obtained from the dielectric spectra
Kremer – Tutorial BDS 2016
Debye Relaxation Havriliak-Negami Relaxation
The dynamics of “water” as seen by BDSThe dynamics of “water” as seen by BDS
BDS - Broadband Dielectric SpectroscopyMaterials with a permanent dipolar momentBDS - Broadband Dielectric SpectroscopyMaterials with a permanent dipolar moment
Cellsf (Hz)f (Hz)HH
HH33NN++ —— C C —— COOCOO--
RR
1010--22
Proteins
Water
1010221010111010--11 101044 101055 101066 101077 101088 101099101033101000 10101010 10101111
Macromolecules
Molecular liquidsPolymer blends
Composites
T = 100-500 K (-173 to 230 oC)
Amino Acids
The dynamics of “water” as seen by BDSThe dynamics of “water” as seen by BDS
BDS - Broadband Dielectric SpectroscopyMaterials with a permanent dipolar momentBDS - Broadband Dielectric SpectroscopyMaterials with a permanent dipolar moment
f (Hz)f (Hz)
1010--22
Water
1010221010111010--11 101044 101055 101066 101077 101088 101099101033101000 10101010 10101111
0.1 1 10 100 10000
20
40
60
80
´,
´´
f [GHz]
T = 293K
Swenson & Bergman, Nature (2001)-2 0 2 4 6
1.5
2.0
2.5
3.0
3.5
4.0
4.5
´´
f [Hz]
T = 165K
0.1 1 10 100 10000
20
40
60
80
´
, ´´
f [GHz]
T = 293K
2 3 4 5 6 7 8-12
-10
-8
-6
-4
-2
0
2
log
()
1000/T [K-1]
Bulk Liquid
Liquid underconfinement
Bulk water - Phase diagram in the No man´s land – EXPERIMENTSBulk water - Phase diagram in the No man´s land – EXPERIMENTS
Outline
‐ Where is water? Several fields of research.
‐ General Phenomenology (glasses, Tg, relaxations) Properties of bulk water / Phase diagram (simulations vs experiments)
-Broadband Dielectric spectroscopy and the study of water molecules
-The dynamics of water at low temperatures in:
- Solution of polymers, glasses and other materials
- Short introduction to Proteins and other biomolecules
- Summary
Calorimetric response of water solutionsCalorimetric response of water solutions
General Result for several materials:
Polymers and small glass formers: 45-50 wt%Sugars: 30-35 wt%
General Result for several materials:
Polymers and small glass formers: 45-50 wt%Sugars: 30-35 wt%
Poly(vinyl methyl ether)
Mixtures from the dry polymer to 50 wt% of water
Mixtures from the dry polymer to 50 wt% of water
Macromolecules (2005)
Cw [wt%] Tg [K]
Dry PVME 0 250.0
PVME-2 2 246.3
PVME-4 4 244.6
PVME-10 15 224.07
PVME-15 10 220.4
PVME-20 20 216.71
PVME-30 30 208.39
PVME-35 35 214.2
PVME-40 40 207.57
PVME-45 45 202.09
PVME-50 50 196.22
150 200 250-10
-5
0
5
10
15
243K
2 wt%
30 wt%
40 wt%
202K
200K
T [K]
Hea
t Flo
w [a
.u.]
Tonset = 190K
50 wt%
PVME water solutions
246K
J. Chem. Phys. (submitted)
Calorimetric response of water solutionsCalorimetric response of water solutions
0 10 20 30 40 50150200250300350400450500
PVME
T g [K]
cw [wt%]
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVP
T g [K]
cw [wt%] 0 10 20 30 40 50150
200
250
300
350
400
450
500 PVP -PLL 3-lysine
T g [K]
cw [wt%]
230 K
50 K
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME 2PG 3PG PPG 5EG
T g [K]
c [wt%]
Plasticization or other mechanism?
The dynamics of dry PVMEThe dynamics of dry PVME
Macromolecules (2005)
10-1 100 101 102 103 104 105 1060.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
280285
275270265260
´´
f [Hz]
PVME Dry 255 K
Now we hydrated the sample with 2 wt% of water…
Tg, dry = 250 K
10-1 100 101 102 103 104 105 1060.00
0.01
0.02
0.03
0.04
0.05
0.06
0.07
PVME Dry
165 170175
160
´´
f [Hz]
155 K
Relaxation Map DRY PVME
3 4 5 6 7-8
-6
-4
-2
0
2
log
( [s
])
1000/T [K-1]
(case Tg,dry – Tg, cw,max < 50 oC)
The dynamics of water in PVME – low water contentThe dynamics of water in PVME – low water content
Macromolecules (2005)
10-1 100 101 102 103 104 105 1060.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
280285
275270265260
´´
f [Hz]
PVME Dry 255 K
10-1 100 101 102 103 104 105 1060.00
0.01
0.02
0.03
0.04
0.05
0.06
0.07
NEW relaxation
´´
f [Hz]
PVME-2cw = 2 wt%
-relaxation
10-1 100 101 102 103 104 105 1060.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
280
285
275270265260
´´
f [Hz]
PVME-2cw = 2 wt%255 K
Tg, dry = 250 K
Tg, 2wt% = 246 K
10-1 100 101 102 103 104 105 1060.00
0.01
0.02
0.03
0.04
0.05
0.06
0.07
PVME Dry
165 170175
160
´´
f [Hz]
155 K
(case Tg,dry – Tg, cw,max < 50 oC)
The dynamics of “water” in PVME – low water contentThe dynamics of “water” in PVME – low water content
At low water content (lower than cw = 5-10 wt%):
- There is a “new” relaxation (water molecules), w (T) is Arrhenius- (T) remains the same increasing water content- (T) faster increasing water content because plasticization
0.22 eV
0.45 eV
3 4 5 6 7-8
-6
-4
-2
0
2 Dry PVME PVME, cw = 2 wt%
log
( [s
])
1000/T [K-1]
3 4 5 6 7-8
-6
-4
-2
0
2 Dry PVME PVME, cw = 2 wt% PVME, cw = 4 wt%
log
( [s
])
1000/T [K-1]
(case Tg,dry – Tg, cw,max < 50 oC)
The dynamics of “water” in PVME – high water contentThe dynamics of “water” in PVME – high water content
10-1 100 101 102 103 104 105 10610-2
10-1
100
101
2
20
10
3035
4045
´
´
f [Hz]
T = 170K50
dry PVME
Macromolecules (2005)
(case Tg,dry – Tg, cw,max < 50 oC)
Relaxation strength and shape factor Relaxation strength and shape factor
0 10 20 30 40 50
0.4
0.5
0.6
01020304050
cw [wt %] General Result:
Water mobility increases around 30 wt% for both polymers and small glasses and 20 wt % for sugars
General Result:
Water mobility increases around 30 wt% for both polymers and small glasses and 20 wt % for sugars
Cole-Cole function
Macromolecules (2005); J NON Crys Solid (2007); PRE (2008); J. Chem. Phys. (2008); PCCP (2010) ; JPCB (2011)
10-1 100 101 102 103 104 105 106
10-1
100
101
220210200190180170160
´´
f [Hz]
PVME - Watercw = 40 wt%
150 K
160 200 240 280
0
5
10
15
20
25
30
35
40
45
50
2
1020
3035
40
45
T [K]
50 wt%
160 200 240 280
0.4
0.5
0.6
0.7
2
T [K]
10
2030
3540
45
50 wt%
(case Tg,dry – Tg, cw,max < 50 oC)
35
30
25
20
15
3 4 5 6 7-12-10
-8-6-4-202
Cp
log
()
1000/ T [K-1]
The dynamics of “water” in PVME – low water contentThe dynamics of “water” in PVME – low water content
10-1 100 101 102 103 104 105 106
10-1
100
101
220210200190180170160
´´
f [Hz]
PVME - Watercw = 40 wt%
150 K
35
30
25
20
15
3 4 5 6 7-12-10
-8-6-4-202
Cp
log
()
1000/ T [K-1]
Tg = 190KPVME
cw = 50wt%
The change from a fragile liquid to strong liquidis produced at Tg
The change from a fragile liquid to strong liquidis produced at Tg
(case Tg,dry – Tg, cw,max < 50 oC)
The dynamics of water in several systems – high water contentThe dynamics of water in several systems – high water content
J. Chem. Phys. (2008)
02468
155K3PG - cw = 50wt%
0
2
4
6
175K
0
2
4
6
´´
188K
Process I
Process II
0369
194K
10-2 10-1 100 101 102 103 104 105 1060
3
6
9
f [Hz]
212K
3-propylene glycol
-2
0
2
4.5 5.0 5.5 6.0 6.5 7.0 7.5-6
-4
-2
0
2
Hea
t flo
w [a
.u.]
Tg = 185 K
log
()
1000/T [K-1]
3PGcw = 40 wt%
The change from a fragile liquid to strong liquidis produced at Tg
The change from a fragile liquid to strong liquidis produced at Tg
(case Tg,dry – Tg, cw,max < 50 oC)
Interpretation of the crossover: Chemical Review (2016)
3 4 5 6 7-12
-9
-6
-3
0
3 5EG PVME glucose 3PG DNA Myoglobin 1PG 2PG 6EG PEG600 Molecular Sieves 3EG 4EG sorbitol
log
(m
ax)
1000/T [K-1]
At high water content
Ea ≈ (0.54 0.04) eV
At high water content
Ea ≈ (0.54 0.04) eV
The dynamics of water in synthetic polymers – high water contentThe dynamics of water in synthetic polymers – high water content
At the highest water concentration (before crystallization)
PRL (2008), PRE (2008)
T < Tg
Local relaxation of water moleculesin soft confinements
Local relaxation of water moleculesin soft confinements
(case Tg,dry – Tg, cw,max < 50 oC)
Comparison solutions at high water content/hard confinementsComparison solutions at high water content/hard confinements (case Tg,dry – Tg, cw,max < 50 oC)
J. Phys.: Cond. Matter (2015)
SOFT confinements: high water concentration (water less influenced by other molecules) HARD confinements: well-defined geometry / uniform filling
3 4 5 6 7 8-12
-10
-8
-6
-4
-2
0
2 PGME (cw = 55 wt%) Mineral Clays Molecular Sieves MCM (C10) Mb (cw = 33 wt%)
log
()
1000/T [K-1]
water in hard confinements
water in solutions
At high water content
Ea ≈ (0.54 0.04) eV
At high water content
Ea ≈ (0.54 0.04) eV
Different scenarios Different scenarios (case Tg,dry – Tg, cw,max < 50 oC)
Chem. Rev., 2016, 116 (13), pp 7608–7625
Summarizing dynamics of water Summarizing dynamics of water
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME
T g [K]
cw [wt%]
50 K
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME 2PG 3PG PPG 5EG
T g [K]
c [wt%]
A single relaxation of water molecules
0 10 20 30 40 50
0
10
20
30
40
50
cw [wt %]
35
30
25
20
15
3 4 5 6 7-12-10
-8-6-4-202
Cp
log
()
1000/ T [K-1]
3 4 5 6 7 8-12
-10
-8
-6
-4
-2
0
2 PVP (cw = 61 wt%) PGME (cw = 55 wt%) Mineral Clays Molecular Sieves MCM (C10) Mb (cw = 33 wt%)
log
()
1000/T [K-1]
(case Tg,dry – Tg, cw,max < 50 oC)
Interpretations: Chem. Rev., 2016, 116 (13), pp 7608–7625
At high water content
Ea ≈ (0.54 0.04) eV
At high water content
Ea ≈ (0.54 0.04) eV
Calorimetric response of water solutionsCalorimetric response of water solutions
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME
T g [K]
cw [wt%]
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVP
T g [K]
cw [wt%]
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVP -PLL 3-lysine
T g [K]
cw [wt%]
230 K
50 K
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME 2PG 3PG PPG 5EG
T g [K]
c [wt%]
Plasticization or other mechanism?
A single relaxation of water molecules
J. Chem. Phys. (submitted)
The dynamics of water – high water contentThe dynamics of water – high water content (case Tg,dry – Tg, cw,max >> 50 oC)
Simultaneous fitting of real and imaginary permittivity
J. Physical Chemistry Letters 7, 4093‐4098 (2016)
100 102 104 1060
10
20
30
10-1 100 101 102 103 104 105 1060.1
1
10
Fast water relaxation
Slow water relaxation
'
Hz
T = 190 K (a)
Slow water relaxation
Fast water relaxation
T = 190 K (b)
''
Hz
10-2 100 102 104 10601020304050
10-2 100 102 104 106
10-1
101
103
Fast water relaxationSlow water
relaxation
(c) T = 207.5 K
'
Hz
Slow water relaxation
(d) T = 207.5 K
''
Hz
Fast water relaxation
100 102 104 106100
101
102
103
100 102 104 10610-1
101
103
105
-relaxation
T = 300 K(e)
'
Hz
(f) T = 300 K
''
Hz
-relaxation
Wübbenhorst and van Turnhout; Journal of Non‐Crystalline Solids 2002, 305, 40‐49
Poly (vinyl pyrrolidone)cw = 40 wt%
The dynamics of water – high water contentThe dynamics of water – high water content (case Tg,dry – Tg, cw,max >> 50 oC)
100 102 104 106
101
100 102 104 10610-1
100
101
10-2 100 102 104 106
101
102
100 102 104 10610-1
101
103
101 104 107
100
102
104
101 104 107100
101
102
103
T = 180 K
'
2
´´
2T = 180 K
2
3'
T = 212 K
´´
T = 212 K
23
f [Hz]´
T = 242 K
43
f [Hz]
43
'
T = 242 K
-Poly(lysine) cw = 40 wt%
Simultaneous fitting of real and imaginary permittivity
J. Physical Chemistry Letters 7, 4093‐4098 (2016)
The dynamics of water – high water contentThe dynamics of water – high water content (case Tg,dry – Tg, cw,max >> 50 oC)
30 35 40 450
30
60
90
120
150
180
205 K 252.5 K 275 K
P3
cw wt%
P4
(a)
30 35 40 4505
101520253035
140K 170K 200K
cw wt%
(b)P2
3 4 5 6 7-12
-9
-6
-3
0
3
3
4
2
3
log
()
1000/T [K-1]
-PLL (cw = 40 wt%)
HF
[a.u
.]
Two relaxations due to water molecules
Two relaxations due to water molecules
J. Physical Chemistry Letters 7, 4093‐4098 (2016)
Processes 3 and 4 are coupled!Processes 3 and 4 are coupled!
Summarizing the response of water in solutionsSummarizing the response of water in solutions
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVP -PLL 3-lysine
T g [K]
cw [wt%]
230 K
50 K
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME 2PG 3PG PPG 5EG
T g [K]
c [wt%]
Plasticization or other mechanism?
A single relaxation of water molecules
A single relaxation of water molecules
Two relaxations of water molecules: the slower one
coupled to the α-relaxation
Two relaxations of water molecules: the slower one
coupled to the α-relaxation
4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5
-6
-4
-2
0
2
1000/T [K-1]
log
()
3PG cw = 50 wt%
Plasticization
3 4 5 6 7-12
-9
-6
-3
0
3
-PLL (cw = 40 wt%)3
4
2
3
log
()
1000/T [K-1]
Outline
‐ Where is water? Several fields of research.
‐ General Phenomenology (glasses, Tg, relaxations) Properties of bulk water / Phase diagram (simulations vs experiments)
-Broadband Dielectric spectroscopy and the study of water molecules
-The dynamics of water at low temperatures in:
- Solution of polymers, glasses and other materials
- Short introduction to Proteins and other biomolecules
- Summary
ProteinsProteins
- Proteins are polymers made out of amino acids
Common backbone
Different side chains Different chemical properties
Amino acids – peptide bondAmino acids – peptide bond
- Proteins are polymers made out of amino acids
Primary structure
Peptides: 2 ‐ 50 amino acids
Proteins : >50 amino acids
ProteinsProteins
Tertiary structureTertiary structure
The function of a protein depends on its tertiary
structure
The function of a protein depends on its tertiary
structure Polymer
Monomer: Amino Acids
Primary structure
-pleated sheet
Secondary structure(Conformations)
-Helix
Dynamics of water in solution of biopolymersDynamics of water in solution of biopolymers
Medium irrelevant! Water solvent play crucial rolesin structure, dynamics and function
Water as a bio-molecule:No passive solvent in biology
Water as a bio-molecule:No passive solvent in biology
Calorimetric response of protein solutionsCalorimetric response of protein solutions
Tg of proteins is no longer observed below ~0.05 wt% of water
0 10 20 30 40 50150
200
250
300
350
400
450
500
0
BSA Gluten Glutenin Gliadin Gelatin Elastin Lysozyme Collagen
T g [K]
cw [wt%]
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVP -PLL 3-lysine
T g [K]
cw [wt%]
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME 2PG 3PG PPG 5EG
T g [K]
c [wt%]
Comparison with soft matter
Collection of different cases
1)Protein in a proper solution (water crystallization)2)Protein in less diluted solution (water crystallization)3)Peptides in solution (no crystallization)
Collection of different cases
1)Protein in a proper solution (water crystallization)2)Protein in less diluted solution (water crystallization)3)Peptides in solution (no crystallization)
Dielectric response of protein in well-diluted solutionsDielectric response of protein in well-diluted solutions
Shinyashiki, N. et al. J. of Phys. Chem. B 113, 14448 (2009)
3 4 5 6 7 8 9 10
-10
-8
-6
-4
-2
0
2
4
6
20BSA-80W
log
1000/T 1/K
IIbIIaIII
IbIa
II: crystallized bulk water -relaxation responsible
for the glass transition
I: uncrystallized water in the
hydration water
- Low solubility degree of proteins- Protein solutions large quantity of water crystallization
Bovine serum albumin (BSA)80 wt% of water – 20 wt% of protein
Dielectric response of protein in more concentrated solutionsDielectric response of protein in more concentrated solutions
3 4 5 6 7 8
-8
-6
-4
-2
0
2
4
40wt%
40wt%28wt%
18wt%
13wt%
process: cooperative motion of protein and water
w process: relaxation of water near the protein surface
ice2-40wt%ice1-40wt%
log
1000/T 1/K
v process: main relaxation of hydration water
cw=7wt%cw=18wt%
28wt%
40wt%cw=28wt%
At low temperatures, two relaxation of water molecules
(similar to “soft matter” with a broad Tg variation with water content)
Dielectric response of protein in more concentrated solutionsDielectric response of protein in more concentrated solutions
Dielectric response of small peptides in solutionDielectric response of small peptides in solution
J Phys. Chem Lett. 7, 4093‐4098 (2016)
3 4 5 6 7-12
-10
-8
-6
-4
-2
0
2
H
(b)
process 1 process 2 process 3 process 4 process 5 cw = 5 wt%
log
()
1000/ T [K-1]3Lys 32Lys
3 4 5 6 7
-8
-6
-4
-2
0
2
3 4 5 6 7-10
-8
-6
-4
-2
0
2
3 4 5 6 7-12
-10
-8
-6
-4
-2
0
2
1000/T [K-1]
3
1
4
2
3
log
()
3-Lys (cw = 40 wt%)
1000/T [K-1]
-helix3
4
2
3
log
()
10-Lys (cw = 40 wt%)
-sheet
3
4
23
log
()
1000/T [K-1]
-PLL (cw = 40 wt%)
Outline
‐ Where is water? Several fields of research.
‐ General Phenomenology (glasses, Tg, relaxations) Properties of bulk water / Phase diagram (simulations vs experiments)
-Broadband Dielectric spectroscopy and the study of water molecules
-The dynamics of water at low temperatures in:
- Solution of polymers, glasses and other materials
- Short introduction to Proteins and other biomolecules
- Summary
Dynamics of ordinary solutions and protein solutions - SUMMARYDynamics of ordinary solutions and protein solutions - SUMMARY
No related dynamics Coupled dynamics Coupled dynamics
SOLUTE SOLVENT ICE
1000/T
Log
()
A single water relaxation
1000/T
Log
()
Two relaxations due to water
1000/T
Log
()
At least two water dynamics
27
cw
TgSolutions of soft matter 1
cw
Tg
Solutions of soft matter 2
cw
Tg
Protein solutions
- Jan Swenson, Chalmers University of Technology
- Gustavo Schwartz, CSIC- Angel Alegria, UPV- Juan Colmenero, UPV- Fabienne Barroso-Bujans, DIPC
- Izaskun Combarro-Palacios, phD student, CFM (Now at Cidetec)- Manuel Monasterio, phD student, CFM (Now at Shenzhen Advanced Civil Engineering Technology, China)- Luciana Saiz, CFM (Now: Researcher in Conicet, Argentina)- Lokendra Singh, Profesor (Now: India)- Jorge Melillo, phD student, CFM
AcknowledgementsAcknowledgements
Travel and local expensessupported by EU
Broadband Dielectric Spectroscopy San Sebastian, SpainBroadband Dielectric Spectroscopy San Sebastian, Spain
THANK THANK YOU YOU FORFOR YOUR YOUR ATTENTIONATTENTION
Amino acid in well-diluted aqueous solutionsAmino acid in well-diluted aqueous solutions
109 10100
10
20
30
40
Water 0.04M 0.11M 0.20 M 0.28 M 0.37 M 0.48 M 0.54 M
Process 2
L-arginine
''
f [Hz]
Process 1
109 10100
20
40
60
80
100
120
Process 2
Process 1
L-arginine
f [Hz]
Water 0.04M 0.11M 0.20 M 0.28 M 0.37 M 0.48 M 0.54 M
'
400 water molecules per AA Well-diluted solutionsWell-diluted solutions
109 10100
20
40
60
80
100
120
10
20
30
40
50
'
''
' ''
L-proline / Water c = 1.07 mol L-1
f [Hz]
'''
Waterrelaxation
Amino acid relaxation
Dynamics of water in solution of biopolymersDynamics of water in solution of biopolymers
Well-diluted solutionsWell-diluted solutions
Bulk-like waterGlobal motion of protein
Origin questioned …
Sokolov et al: This relaxation is mainly due to protein atoms and hydration water relaxes at much shorter time scales
S. Khodadadi et al, J. Phys. Chem. B, 2011, S. Khodadadi,et al, J. Chem. Phys., 2008
Where is found water? – Geological water – Confined waterWhere is found water? – Geological water – Confined water
Size of confinements:- micro, d < 20 Å- meso, 20 Å < d < 500 Å; - macro, d > 500 Å
Vermiculite Clays
Swenson‐Bergman, Nature, 2001
Cerveny et al, JCP (2011)
MCM 41
21 Å or 36 ÅSjöström et al, JCP 128, 154503 (2008)
12 Å
OXIDATION
Graphite Oxide
Increasing hydration
8Ǻ6Ǻ
The dynamics of water in synthetic polymers – high water contentThe dynamics of water in synthetic polymers – high water content
10-1 100 101 102 103 104 105 10610-2
10-1
100
101
2
20
10
3035
4045
´´
f [Hz]
T = 170K50
dry PVME
Deuterated water is slower than protonated water
100 101 102 103 104 105 106 1070.1
1
10
deuterated sample protonated sample
´´
f [Hz]
Water molecules are responsible for this “new” dielectric process
Crossover strong-to-fragile? Crossover strong-to-fragile?
-2
0
2
4.5 5.0 5.5 6.0 6.5 7.0 7.5-6
-4
-2
0
2
Hea
t flo
w [a
.u.]
Tg = 185 K
log
()
1000/T [K-1]
3PGcw = 40 wt%
Chen et al PNAS (2006)Mallamace et al JCP (2007)Chu et al PRE (2008)…..
“Confined Waters exhibit a LL transition at 225 K”
“Fragile to strong transition”
“the low temperature Arrhenius process is most likely of local character”
Chen et al, PRE 2014
“the low temperature Arrhenius process is most likely of local character”
Chen et al, PRE 2014
The crossover does not represents the fragile-to-strong transition of BULK water
The crossover does not represents the fragile-to-strong transition of BULK water
Chemical Review (2016)
Resume results of dynamics of water in synthetic materialsResume results of dynamics of water in synthetic materials
Low water content (water influenced by the solute)
Low water content (water influenced by the solute)
High water content (clusters of water less influenced by the solute)
High water content (clusters of water less influenced by the solute)
Solutions of water and polymers, sugars, glass forming materials…
2
1000/T
Log
()
Tg,DSC
2
1000/T
Log
()
Tg,DSC
cw
cw
Water mobility increases around (c*):
30 wt% for both polymers and small glasses
20 wt % for sugars
Water mobility increases around (c*):
30 wt% for both polymers and small glasses
20 wt % for sugars
Ea = (0.54 ± 0.04) eV
c* c*
SummarizingSummarizing
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME
T g [K]
cw [wt%]
50 K
0 10 20 30 40 50150
200
250
300
350
400
450
500 PVME 2PG 3PG PPG 5EG
T g [K]
c [wt%]
Low water content
(water influenced by the solute)
Low water content
(water influenced by the solute)
High water content (clusters of water less
influenced by the solute)
High water content (clusters of water less
influenced by the solute)
2
1000/T
Log
()
Tg,DSC
Water mobility increases at c*Water mobility increases at c*
2
1000/T
Log
()
Tg,DSC
Ea = 0.54 eV
cw
c*
case Tg,dry – Tg, cw,max < 50 oC
Water in hard confinementsWater in hard confinements
L
L
pores: 2-D confinementlayers: 1-D confinement
Well-defined confinement systems (HARD CONFINEMENT)
MCM-41: Swenson et al, Chen et al, Mallamace et al, Vogel et al…
Cement-like materials: Shinyashiki, Fratini et al, Cerveny et al…
Graphite Oxide: Cerveny et al
Minerals and Clays: Swenson et al, Feldman et al, Bruni et al,, Maheshwari et al…
Hydrogels: Bruni et al, Pissis et al, Shinyashiki at al…
Molecular Sieves: Swenson et al, Bruni et al, Oguni et al…
Disordered systems: 3-D confinement
water in hard confinementswater in hard confinements
OXIDATION
Graphite Graphite Oxide
Graphite OxideGraphite Oxide
Increasing hydration
8Ǻ6Ǻ
J. Phys. Chem C (2010)
MCM 41MCM 41
The pores are ordered in a hexagonal structure
The pore diameter is d = 2.1 nm
Sjostrom, Swenson, et al, J. Chem Phys (2008)
100 150 200 250 300 350
-0.2
0.0
0.2
heating
25 wt% dry GO
Rev
. Hea
t Flo
w [a
.u.]
T [K]
cooling
In hard confinements: -No crystallization - No observation of a glass transition by DSC
In hard confinements: -No crystallization - No observation of a glass transition by DSC
Water in hard confinementsWater in hard confinements
4 6 8-12-10-8-6-4-202
1000/T [K-1]
log()
5% 10% 15% 17% 20% 25%
No dependence on water contentNo dependence on water content
10-2 100 102 104 106 108
0.1
1
160
170
180
200
225K
150140
´´
f [Hz]
130
Water in Graphite Oxide
3 4 5 6 7 8-12
-10
-8
-6
-4
-2
0 GO - NS GO - cw = 25 wt% - BDS
log
()
1000/T [K-1]
Water in Graphite Oxide
4 6 8
-10
-8
-6
-4
-2
0
2
Dry 14 wt% 28 wt% 111 wt%
1000/T [K-1]
log
()
MCM41
2 4 6 8-12
-10
-8
-6
-4
-2
0
2
GO-25wt% Molecular Sieves Silica hydrogel Mineral Clays MCM (C10) C-S-H
1000/T [K-1]
log
()
Water in hard confinementsWater in hard confinements
J. Phys.: Cond. Matter (2015)
Above the crossover – Both the time scale and Ea are differentfor all types of confinements
Below crossover – Ea is similar for all confinements
Ea ≈ (0.54 0.04) eV
-rela
xation
of wate
r mole
cules
in hard
confi
nement
s
-rela
xation
of wate
r mole
cules
in hard
confi
nement
s
Differences and Similarities of water in soft and hard confinementsDifferences and Similarities of water in soft and hard confinements
J. Phys.: Cond. Matter (2015)
SOFT confinements: high water concentration (water less influenced by other molecules) HARD confinements: well-defined geometry / uniform filling
3 4 5 6 7 8-12
-10
-8
-6
-4
-2
0
2 PVP (cw = 61 wt%) PGME (cw = 55 wt%) Mineral Clays Molecular Sieves MCM (C10) Mb (cw = 33 wt%)
log
()
1000/T [K-1]
water in hard confinements
water in solutions
2 4 6 8-12-10-8-6-4-202
GO-25wt% - BDS GO - NS
log()
1000/T [K-1]
Summarizing - Water in soft and hard confinementsSummarizing - Water in soft and hard confinements
No calorimetric “Tg”
-8
-6
-4
4.5 5.0 5.5 6.0 6.5 7.0
-6
-4
-2
0
2
cw = 40 wt%5EG
log
()
1000/T [K-1]
Tg, DSC = 176K
HF
[a.u
.]
Soft confinements Hard Confinements
The change in the dynamics is produced in the temperature range (150-200) K depending on the
confinement system
The change in the water dynamics is produced at Tg.
4 5 6 7-8
-6
-4
-2
0
2
50% 40% 30% 20%
log
()
1000/T [K-1]
cw
Soft Confinements
SOFT CONFINEMENTS
Calorimetric glass transition(solvent + solute)
+
Stronger concentration dependence
SOFT CONFINEMENTS
Calorimetric glass transition(solvent + solute)
+
Stronger concentration dependence
3PG-water
HARD confinements
HARD CONFINEMENTS
No calorimetric glass transition (exception Oguni et al (2011) Tg = 210 K)
+
Almost no concentration dependence
HARD CONFINEMENTS
No calorimetric glass transition (exception Oguni et al (2011) Tg = 210 K)
+
Almost no concentration dependence
4 6 8
-10
-8
-6
-4
-2
0
2
Dry 14 wt% 28 wt% 111 wt%
1000/T [K-1]
log
()
MCM41
Summarizing - Water in soft and hard confinementsSummarizing - Water in soft and hard confinements
J. Phys.: Cond. Matter (2015)
Origin of the crossover (hard and soft confinements)Origin of the crossover (hard and soft confinements)
“Cooperative re-arrangement regions” (CRR)
4 5 6 7 8 9 10 11
-8
-6
-4
-2
0
2
log
()
1000/T [K-1]
4 5 6 7 8 9-8
-6
-4
-2
0
log
()
1000/T [K-1]
Introduction - Protein dynamicsIntroduction - Protein dynamics
If nothing can move, nothing can functionIf nothing can move, nothing can function
Perutz (1970): described the movements hemoglobin must undergo to fulfill its function
Frauenfelder (1985): revealed a hierarchical organization of proteins motions
Perutz Nature 228, (1970)P. W. Fenimore, H. Frauenfelder et al., PNAS 110, 14408 (2004)
A protein does not exist in a unique conformation
A protein can assume a very large number of somewhat different conformations
Ec
ccEc
cc0Ec
cc1Ec
cc2
State
Tier0
Tier1
Tier2
A0 A1 A3
log
()
1/T
log
()
1/T
“solvent-slaved” motions“slaving” behaviour “solvent-slaved” motions“slaving” behaviour
Fluctuations of the protein (which follows aVFT behaviour) come from the solvent
Myoglobin: escape of CO from the interior of the protein
P. W. Fenimore et al., PNAS 110, 14408 (2004)
3 4 5 6 7 8-12
-10
-8
-6
-4
-2
0
exit
HF
[a.u
.]
1000/T [K-1]
log
()
2.5 3.0 3.5 4.0 4.5 5.0 5.5
-8
-6
-4
-2
0
2
3 Lysine-dry
log
()
1000/T [K-1]2.5 3.0 3.5 4.0 4.5 5.0 5.5
-8
-6
-4
-2
0
2
3 Lysine-dry cw < 1wt%
log
()
1000/T [K-1]
3-Lysine cw = 0 wt% cw = 40 wt%
2.5 3.0 3.5 4.0 4.5 5.0 5.5
-8
-6
-4
-2
0
2 cw = 10 wt%
3 Lysine-dry cw < 1wt%
log
()
1000/T [K-1]2.5 3.0 3.5 4.0 4.5 5.0 5.5
-8
-6
-4
-2
0
2 cw = 10 wt% cw = 40 wt%
3 Lysine-dry cw < 1wt%
log
()
1000/T [K-1]
The dynamics of water – From low to high water contentThe dynamics of water – From low to high water content (case Tg,dry – Tg, cw,max >> 50 oC)
Relaxations are coupled at water content higher than 15 wt%
At low water content, a single relaxation
These findings further corroborate the existence of a strong coupling between protein fluctuations and solvent dynamics.
Our results indicate a slowdown of solvent dynamics in the immediate vicinity of the protein. Hence, there is a mutual influence of protein
and solvent dynamics, rather than a full slaving.
“protein-solvent couplings can be different for side chain and backbone units”
The hydration dynamics change depending on the different DNA exposed sites. The waters confined in the narrow minor
groove are much more significantly retarded than in bulk and therefore it seems that the biomolecule slaves the water
which is the opposite of the slaving picture.
Introduction –Slaving or not?Introduction –Slaving or not?
“The hydration water dynamics is always faster than protein side-chain relaxations but with the same energy barriers, indicating that the
hydration shell fluctuations driving protein side-chain motions on the picosecond time scales and thus elucidating their ultimate relationship”
Bulk water - Phase diagram in the No man´s landBulk water - Phase diagram in the No man´s land
Stanley, 1992Stanley, 1992
Below -75 oC… they found a low density liquid and a high density liquid: LDL and HDL.
Liquid-liquid critical point hypothesis
Bulk water - Phase diagram in the No man´s land - SIMULATIONSBulk water - Phase diagram in the No man´s land - SIMULATIONS
Stanley, 1992Stanley, 1992
Limmer-Chandler, 2011-2013Limmer-Chandler, 2011-2013
“It is not enough to wait for the density of the system to settle down to a steady state”
The problem is the equilibration time!!!!
Debenedetti,2013Debenedetti,2013
He remains convinced that the liquid–liquid transition is real
"Our calculations are completely inconsistent with David Chandler's – we clearly see two, not one, liquid phases."
Below -75 oC… they found a low density liquid and a high density liquid: LDL and HDL.
Liquid-liquid critical point hypothesis
The dynamics of water – high water contentThe dynamics of water – high water content (case Tg,dry – Tg, cw,max >> 50 oC)
3 4 5 6 7-12
-9
-6
-3
0
3
3
4
2
3
log
()
1000/T [K-1]
-PLL (cw = 40 wt%)
HF
[a.u
.]
3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5-8-7-6-5-4-3-2-1012
cw=45wt%cw=40wt%cw=35wt%cw=30wt%
log
(
1000/T 1/K
PLLProcess2
3.0 3.5 4.0 4.5 5.0 5.5 6.0
-10
-8
-6
-4
-2
0
2
45 wt% 40 wt% 35 wt% 30 wt%
log
(3)
1000/T [K-1]
-PLLProcess 3
3.0 3.5 4.0 4.5 5.0-8
-6
-4
-2
0
2
-PLLProcess 4
45 wt% 40 wt% 35 wt% 30 wt%
log
(4)
1000 /T [K-1]
cwProcesses 3 and 4 are coupled!Processes 3 and 4 are coupled!
J. Physical Chemistry Letters 7, 4093‐4098 (2016)