CHAPTER 2

Water and Aqueous Solutions

Types of non-covalent interactions between molecules
Properties of water THE medium for life
Hydrophobic -- nonpolar -- moieties aggregate in water
Solute effects on bulk properties of water
Weak acids and bases
Buffers theory and practice
Water as participant in biochemical reactions
Learning Objectives
Physics of Non-covalent Interactions

Ionic (Coulombic) Interactions
Electrostatic interactions between permanently charged species,
or between the ion and a permanent dipole
Dipole Interactions
Electrostatic interactions between uncharged, but polar molecules
Van der Waals Interactions
Weak interactions between all atoms, regardless of polarity
Attractive (dispersion) and repulsive (steric) component

Hydrophobic Effect
Complex phenomenon associated with the ordering of water molecules around
non-polar substances
Non-covalent interactions do not involve sharing a pair
of electrons. Based on their physical origin, one can
distinguish between
Noncovalent
Forces and
Interactions
Hydrogen bonds
Ion-Ion 1/r
Ion-dipole 1/r
2
Dipole-dipole 1/r
3
Dipole - Induced
dipole - 1/r
5

ID ID (Van der
Waals) - 1/r
6
Hydrophobic

Hydrogen Bonds
Strong dipole-dipole or charge-dipole interaction that arises
between an acid (proton donor) and a base (proton acceptor)
Typically 4-6 kJ/mol for bonds with neutral atoms,
and 6-10 kJ/mol for bonds with one charged atom
Typically involves two electronegative atoms (frequently
nitrogen and oxygen)
Hydrogen bonds are strongest when
the bonded molecules are oriented to
maximize electrostatic interaction.
Ideally the three atoms involved are in a line
Hydrogen
Bonds:
Importance of Hydrogen
Bonds
Source of unique properties of water
Structure and function of proteins
Structure and function of DNA
Structure and function of polysaccharides
Binding of a substrates to enzymes
Binding of hormones to receptors
Matching of mRNA and tRNA
Biological Relevance of
Hydrogen Bonds
Van der Waals Interactions
Van der Waals interactions
have two components:
Attractive force (London
dispersion) Depends on
the polarizability
Repulsive force (Steric
repulsion) Depends on
the size of atoms
Attraction dominates at
longer distances (typically
0.4-0.7 nm)
Repulsion dominates at very
short distances
There is a minimum energy
distance (van der Waals
contact distance)
Biochemical Significance of
Van der Waals Interactions
Weak individually
Easily broken, reversible
Universal:
Occur between any two atoms that are near each other
Importance
determines steric complementarity
stabilizes biological macromolecules (stacking in DNA)
facilitates binding of polarizable ligands
Water is the Medium for Life
Life evolved in water
(UV protection)
Organisms typically
contain 70-90% water
Chemical reactions
occur in aqueous
milieu
Water is a critical
determinant of the
structure and function
of proteins, nucleic
acids, and membranes
Structure of the Water
Molecule
Four electron pairs on four
sp
3
orbitals (distorted
tetrahedron)
Two pairs covalently link
hydrogen atoms to a central
oxygen atom.
Two remaining pairs remain
nonbonding (lone pairs)
The electronegativity of the
oxygen atom induces a net
dipole moment
Water can serve as both a
hydrogen bond donor and
acceptor.
Hydrogen Bonding in Water
Up to four H-bonds H
2
O
high boiling point
high melting point
large surface tension
Hydrogen bonding in water
is cooperative.
Hydrogen bonds between
neighboring molecules are
weak (20 kJ/mole) relative
to the HO covalent bonds
(420 kJ/mol)
Water as a Solvent

Water is a poor solvent for nonpolar
substances
nonpolar gases
aromatic moieties
aliphatic chains


Water is a good solvent for charged and
polar substances
amino acids and peptides
small alcohols
carbohydrates

Water Dissolves Many Salts
High dielectric constant of
water ( = 80 ) shields
oppositely charged ions;
Almost no attraction > 40 nm
Electrostatics of solvation
lowers the energy of the
system
Entropy increases as
ordered crystal lattice is
dissolved

NaCl(s) <=>Na
+
+ Cl
-
Ice: H
2
O(s)
Water has many different
crystal forms; the hexagonal ice
is the most common

Hexagonal ice forms a regular
lattice, and thus has a low
entropy

Hexagonal ice has lower
density than liquid water; ice
floats
The Hydrophobic Effect
Refers to the association or folding of non-
polar molecules in the aqueous solution
Is one of the main factors behind:
Protein folding
Protein-protein association
Formation of lipid micelles
Binding of steroid hormones to their receptors
Does not arise because of some attractive
direct force between two non-polar molecules

Solubility of Polar and Non-
polar Solutes
Why are non-polar molecules poorly soluble in water?
Low Solubility of Hydrophobic Solutes
Disruption of H-
bonded H
2
O
networks

Ordered Water
near a hydrophobic
solute

Cavity formation in
a medium with high
surface tension
Hydrophobic Effect
Lipid molecules disperse in the
solution; nonpolar tail of each lipid
molecule is surrounded by ordered
water molecules
Lipid aggregates Water released,
surface area reduced

Hydrophobic Effect Favors
Ligand Binding
Binding sites in
enzymes and receptors
are often hydrophobic
Such sites can bind
hydrophobic substrates
and ligands such as
steroid hormones
Many drugs are
designed to take
advantage of the
hydrophobic effect

Colligative Properties
Some properties of solution boiling point,
melting point, and osmolarity do not depend
strongly on the nature of the dissolved
substance. These are called colligative
properties
Other properties viscosity, surface tension,
taste, and color, among other depend
strongly on the chemical nature of the solute.
These are non-colligative properties.
Cytoplasm of cells are highly concentrated
solutions and have high osmotic pressure

Effect of
Extracellular
Osmolarity
Osmotic Pressure
For a single solute

= RT (ic)

Where i is extent of
dissociation and c is
concentration.

For mixtures
= RT (ic)



Bound Water in Proteins
Ionization of Water
O-H bonds are polar and can dissociate heterolytically
Products are a proton (H
+
) and a hydroxide ion (OH
-
)
Dissociation of water is a rapid reversible process
Most water molecules remain un-ionized, thus pure water
has very low electrical conductivity (resistance: 18 MOcm)
The equilibrium H
2
O H
+
+ OH
-
is strongly to the left
Extent of dissociation depends on the temperature

H
2
O H
+
+ OH
-

Proton Hydration
Protons do not exist free in solution.
They are immediately hydrated to form hydronium
(oxonium) ions
A hydronium ion is a water molecule with a proton
associated with one of the non-bonding electron pairs
Hydronium ions are solvated by nearby water molecules
The covalent and hydrogen bonds are interchangeable.
This allows for an extremely fast mobility of protons in
water via proton hopping
Proton
Hopping
Hydrogen bonded networks
form natural chains for rapid
Proton transfer
Ionization of Water:
Quantitative Treatment
Concentrations of participating species in an equilibrium process
are not independent but are related via the equilibrium constant


H
2
O H
+
+ OH
-
K
eq
=
[H
+
][OH
-
]
[H
2
O]
K
eq
can be determined experimentally, it is 1.810
-16
M at 25 C
[H
2
O] can be determined from water density, it is 55.5 M

Ionic product of water:


In pure water [H
+
] = [OH
-
] = 10
-7
M
2 14 -
2
M 10 1 ] OH ][ H [ ] O H [
+
= = =
eq w
K K
What is pH?

pH is defined as the negative
logarithm of the hydrogen ion
concentration.

Simplifies equations
The pH and pOH must always
add to 14

pH can be negative ([H
+
] = 6 M)
In neutral solution, [H
+
] = [OH
-
]
and the pH is 7
pH = -log[H
+
]
2 14 -
M 10 1 ] OH ][ H [
+
= =
w
K
14 ] OH log[ ] H log[
-
+ =
+
14 pOH pH = +
pH Scale: 1
unit = 10-fold
Dissociation of Weak Electrolytes:
Principle
Weak electrolytes
dissociate only partially
in water
Extent of dissociation is
determined by the acid
dissociation constant K
a

We can calculate the
pH if the K
a
is known.
But some algebra is
needed!
C H
3
O
OH
C H
3
O
O
+
H
2
O
-
+
H
3
O
+
K
eq
] O H [
2
=
eq a
K K
M 10 74 . 1
COOH] CH [
] COO CH ][ H [
5
3
-
3
+
= =
a
K
] COO CH [
] COOH CH [
] [
3
3

+
=
a
K H
Dissociation of Weak Electrolytes:
Example
What is the final pH of a solution when 0.1 moles
of acetic acid is adjusted to 1 L of water?
C H
3
O
OH
C H
3
O
O
-
+
H
+
K
a
0.1 x x x
M 10 74 . 1
x] - 0.1 [
] x ][ x [
5
= =
a
K
x 10 74 . 1 10 74 . 1 x
5 6 2
=
0 10 74 . 1 x 10 74 . 1 x
6 5 2
= +

We assume that the
only source of H
+
is
the weak acid
To find the [H
+
], a
quadratic equation
must be solved.
Ax
2
+ bx + c = 0
Dissociation of Weak Electrolytes:
Simplification
C H
3
O
OH
C H
3
O
O
-
+
H
+
K
a
0.1 x x x
0.1 x x
M 10 74 . 1
0.1] [
] x ][ x [
5
= =
a
K
6 2
10 74 . 1 x

=
x = 0.00132, pH = 2.880
The equation can be
simplified if the amount of
dissociated species is much
less than the amount of
undissociated acid

Approximation works for
sufficiently weak acids and
bases

Check that x << [Total Acid]
pK
a
measures acidity
pK
a
= -log K
a
(strong acid large K
a
small pK
a
)
Buffers are mixtures of weak
acids and their anions
Buffers resist change in pH

At pH = pK
a
, there is a 50:50 mixture of acid and
anion forms of the compound

Buffering capacity of acid/anion system is
greatest
at pH = pK
a

Buffering capacity is lost when the pH differs from
pK
a

by more than 1 pH unit
HendersonHasselbalch Equation:Derivation
Biological Buffer Systems
Maintenance of intracellular pH is vital to all cells
Enzyme-catalyzed reactions have optimal pH
Solubility of polar molecules depends on H-bond donors and acceptors
Equilibrium between CO
2
gas and dissolved HCO
3
-
depends on pH

Buffer systems in vivo are mainly based on
phosphate, concentration in millimolar range
bicarbonate, important for blood plasma
histidine, efficient buffer at neutral pH

Buffer systems in vitro are often based on sulfonic acids of cyclic amines
HEPES
PIPES
CHES


N N
O H
SO
3
Na
Water as a reactant in biochemistry
Chapter 2: Summary
The goal of this chapter was to help you to better
understand:

The nature of intermolecular forces
The properties and structure of liquid water
The behavior of weak acids and bases in water
The way water can participate in biochemical
reactions