Lecture 13

Multiplicity and statistical

definition of entropy

Readings:

Lecture 13, today: Chapter 7: 7.1 – 7.19

Lecture 14, Monday: Chapter 7: 7.20 - end
MCB65
2/26/16 1
 Today’s Goals

Concept of entropy from statistical thermodynamics

Bulk properties of large collections of events/molecules
Multiplicity
Macro and microstates

Statistical definition of the second law of thermodynamics

For a spontaneous process, the multiplicity of the system
increases
Statistical treatment of multiplicity explains macroscopic
properties – back to the ideal gas expansion
MCB65
2/26/16 2
 Hemoglobin and probability
Oxygen binding molecule. Its quaternary structure is a
tetramer composed of two α- and β- subunits.
Assume that for each heme:

p(bound)=1/2

p(unbound)=1/2

MCB65
2/26/16 3
 Independent events

The probability of two

independent events is the
½ ½ ½ ½ product of their individual
probabilities.

p = ½ x ½ x ½ x ½ = 1/16
½ ½ ½ ½

Therefore, for each

hemoglobin, the probability of
obtaining any given outcome is
½ ½ ½ ½ 1/16
MCB65
2/26/16 4
Heme position
1 2 3 4 Multiplicity

What if all you care about is the number of

heme groups that either have or do not have
oxygen bound?

Aggregate the relevant possible outcomes

together
# of desired outcomes
p
total # of possible outcomes

Number of desired outcomes is the

multiplicity, W
MCB65
2/26/16 5
 Multiplicity of a molecular system

The multiplicity of a molecular system is the number of

different molecular configurations consistent with the
macroscopic parameters (e.g. temperature, volume,
concentration) that define the system

A microstate is one particular configuration of molecules

that is consistent with the global macroscopic parameters
that define a particular state of the system

The multiplicity is the number of microstates

MCB65
2/26/16 6
Heme position
1 2 3 4 Multiplicity
4 bound & 0 unbound → W = 1 → p = 1/16 = 0.0625

3 bound & 1 unbound → W = 4 → p = 4/16 = 0.25

2 bound & 2 unbound → W = 6 → p = 6/16 = 0.375

1 bound & 3 unbound → W = 4 → p = 4/16 = 0.25

0 bound & 4 unbound → W = 1 → p = 1/16 =0.0625

MCB65
2/26/16 7
 How to compute multiplicity

When we get to a large number of outcomes / microstates,

it gets tedious to classify and count all of these

For the binomial case (i.e. two possible outcomes, such as

bound/unbound), we can generalize this as:
M!
W
N!( M  N )!

Where N is the number of bound hemes obtained in a total

of M hemes
MCB65
2/26/16 8
 Calculating multiplicity using factorials
Calculate W for 2 bound & 2 unbound in a hemoglobin:

Calculate W for 4 bound& 0 unbound in a hemoglobin:

Note: 0! = 1 (the product of no numbers at all is 1)

MCB65
2/26/16 9
Ways of binding oxygen
Four hemes, labeled 1
through 4
(numbers represented by
different colors)

In how many possible

ways can the hemes
be chosen?

4 choices x
3 choices x
2 choices x
1 choice
4!=24 MCB65
2/26/16 10
 Multiplicity of 2 bound & 2 unbound

2
crimson
bound
sites
2 blue
unbound
sites

MCB65
2/26/16 11
 Multiplicity of 2 bound & 2 unbound
In the 24 outcomes, there are
several instances of different
outcomes with the same
2 crimson bound/unbound pattern
bound We’re over-counting these
sites instances
2 blue
unbound
sites By how much?
There are 2! ways of rearranging
the bound sites and 2! ways of
rearranging the unbound sites
W = 4!/(2! x 2!) = 6

MCB65
2/26/16 12
 Hemoglobin multiplicity
M!
Outcome Example W
N!( M  N )!
4!
4 bound & no unbound W 1
4!0!
4!
3 bound & 1 unbound W 4
3!1!
4!
2 bound & 2 unbound W 6
2!2!
4!
1 bound & 3 unbound W 4
1!3!
4!
0 bound & 4 unbound W 1
0!4!

MCB65
2/26/16 13
 Over-counting in a bigger dataset
2 hemoglobins, 8 hemes, 5 bound and 3 unbound
Number of ways to pick the 8 sites in random order: 8!
5 3 4 2 1 2 3 1
Two microstates
of the same
outcome
1 1 3 4 2 3 5 2

1 3 4 5 2

Ways to arrange 5 bound states: 5 x 4 x 3 x 2 x 1 = 120 = 5!

Same for the 3 unbound states: 3!
Over-counting for this red-blue pattern: 5! x 3!
Multiplicity = 8!/5!3! = M!/(N!(M-N)!)
Where M = # hemes, N = # bound states and (M-N) = # unbound states
MCB65
2/26/16 14
 As M increases…
Multiplicity (W) 4 Hemes (1 Hemoglobin) 10 Hemes

Multiplicity (W)
Bound O2 molecules Bound O2 molecules
200 Hemes 1000 Hemes
Multiplicity (W)

Multiplicity (W)

Bound O2 molecules Bound O2 molecules

MCB65
Modified from The Molecules of Life (© Garland Science 2008) 2/26/16 15
 250 hemoglobins-1000 hemes
M = 1000, N = 500 oxygen-bound hemes
The multiplicity of this outcome is:

1000!
W
500!500!

Stirling’s approximation is a useful mathematical tool to

calculate values for large factorials
ln n! n ln n  n
Where n is a large number
MCB65
2/26/16 16
 Using Stirling’s approximation
M = 1000, N = 500 oxygen-bound hemes 1000!
W
500!500!

Applying a natural log to both sides: ln W  ln 1000!

500!500!
Then: ln W  ln(1000!)  ln(500!)  ln(500!)
 ln(1000!)  2 ln(500!)

Applying Stirling’s approximation: ln n! n ln n  n

ln W  1000 ln 1000  1000
 2(500 ln 500  500)

W = e694 ~ 10301 MCB65

2/26/16 17
 Probability of N=500?

W500 ~ 10301

Number of possible outcomes: 21000 ~ 10301

p500 = multiplicity / # of possible outcomes

~ 10301/10301 ~ 1

Essentially all sequences of 1000 will contain an equal

number of bound and unbound hemoglobin proteins

MCB65
2/26/16 18
Other outcomes still have (vanishingly small) probability

Important to keep in mind that the probabilities are still a

distribution
1000 Hemes

Multiplicity (W)

Bound O2 molecules

For 1000 hemes (250 hemoglobin molecules),

p(N=0) ~ 1/10301
MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 19
 Probability and multiplicity

The relative probability of two outcomes is the ratio of

their respective probabilities
Example, 1000 hemes, N=500 vs. N=600

p500 W500 21000 W500

 1000

p600 W600 2 W600
It’s also the ratio of their multiplicity!
In our example: ln (W500/W600) = 694 – 674 = 20
W500/W600 = e20 = ~5x108

Again, an equal number of bound and unbound is by far

the most likely outcome MCB65
2/26/16 20
 Maximal multiplicity when the derivative is 0
How can we find the maximal multiplicity?
When the slope or derivative of the function is 0 and
(N ≠ 0) or (N ≠ M)

That is: W = f(N) and dW/dN = 0 is a condition for

maximal multiplicity

MCB65
2/26/16 21
 W and lnW have the same maximum

Bound O2 molecules

lnW

So is dlnW/dN = 0

Stirling’s approximation
will be extremely helpful Bound O2 molecules
MCB65
Modified from The Molecules of Life (© Garland Science 2008) 2/26/16 22
 Maximal multiplicity
How can we find the maximal multiplicity?

For the binomial case, it can be derived, using Stirling’s

approximation, that:
d ln W
  ln N  ln( M  N )
dN

At a maximum: d lnW/dN = 0, such that

lnN = ln(M-N)
N = M-N
2N = M
M = N/2
MCB65
2/26/16 23
 Maximal multiplicity leads to maximal entropy
What we’ve learned from hemoglobin:
The observed outcome of an experiment involving a large number
of trials is the one that corresponds to maximum multiplicity

This is broadly applicable beyond hemoglobin

Hopefully, it’s starting to sound like the 2nd law of

thermodynamics!

Much of thermodynamics is about finding the conditions where

multiplicity is maximal

MCB65
2/26/16 24
 Back to molecules: starting with an ideal gas

Isothermal expansion example from Lecture 9

Here dU = dq + dw = 0 because T = constant

Therefore, dq = - dw

Even though dU = 0, there is a change in the system

We stated that “entropy” drives the change

MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 25
 Back to molecules: starting with an ideal gas

Let’s now consider that this compressed gas is in fact made

of particles, gas molecules
More space is available to these molecules when the gas expands
What happens to the multiplicity of the system?
MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 26
 Multiplicity in a molecular system
The multiplicity of a molecular system is defined as the
number of different configurations or conformations of the
component particles that are equivalent

Note: in reality, a
configuration includes the
description of kinetic energy
of each molecule. Here, we’ll
focus only on the position
component

Ideal gas: particles have positions in the system volume

Grid boxes of arbitrary size, small enough to uniquely define the
position of the atoms
MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 27
 Calculating multiplicity

There are 49! ways to put 49 particles in 49 boxes

MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 28
 6 particles in a box
Similar to the case of bound and unbound hemes:
M = number of boxes
N = number of red particles
(M – N) = number of empty boxes (“blue particles”)
M! 49!
W   1.4 107
N!( M  N )! 6!(49  6)!

MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 29
 Multiplicity increases as volume increases

Double the volume of the box and the multiplicity is now:

98!
W
6!(98  6)!
ln W  ln 98! ln 6! ln 92!
 98 ln 98  98  ln 720  (92 ln 92  92)
 20.72
W  109
Figure from
MCB65
The Molecules of Life
2/26/16 30
(© Garland Science 2008)
 Atoms are likely to spread
The two configurations shown below are equally likely in a
larger space

But from our calculations, there are ~1.4x107

configurations equivalent to the one on the left, and ~109
configurations to the one on the right

We’re therefore ~100 times more likely to see the atoms

on both sides of the box MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 31
 Equilibrium state of a system
A microstate is a particular configuration of the atoms or
molecules in a system
Instantaneous snapshot
A state is a global description of the system
Uses bulk properties like temperature, pressure, number of
molecules
The multiplicity of a state is the number of corresponding
microstates
The higher the multiplicity, the higher the likelihood of
observing that state
The system will spontaneously evolve towards the state with
highest multiplicity MCB65
2/26/16 32
Maximal multiplicity defines equilibrium state
Sudden increase in volume
of the system

Over time, the atoms

spread to increase
multiplicity, until they are
evenly distributed, which
has maximal multiplicity

The multiplicity of the

system drives the
expansion
MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 33
If we combine systems A and B, what would the
combined W be?

MCB65
2/26/16 34
WA+B is the product of WA and WB

MCB65
Figure from The Molecules of Life (© Garland Science 2008) 2/26/16 35
lnW is an additive and extensive property
Additive
System A System B

WA WB W=WA×WB
ln W = ln WA + ln WB

Extensive
System 1 System 2

W=(W1)2
W=W1 2x lnW=2ln(W1)

MCB65
2/26/16 36
 Use of lnW for large systems

W rapidly becomes unmanageably large, lnW increases

slower with the # of particles

Notice lnW is an extensive and additive property of the

system

Combining systems A and B to yield (A+B)

WA+B = WA x WB
lnWA+B = lnWA + lnWB

lnW is a state function, it depends only on the parameters

of the present state of the system
MCB65
2/26/16 37
 Statistical definition of entropy

Entropy is defined as:

S  kB ln W
Where kB is the Boltzmann constant
kB = 1.38 x 10-23 JK-1
= R/NA = 8.314 JK-1/6.023 x 10-23

Entropy is a state function of the system

MCB65
2/26/16 38
 Some concepts to remember

Multiplicity (W) is the number of microstates for a given

conformation of the system

A system will spontaneously evolve towards the state of

maximal multiplicity

Entropy is a function of multiplicity

Additive property
Extensive property
State function

MCB65
2/26/16 39