PHY332 Assignment #5 Tim Henley 1000234506

Problem 2

In order to solve this problem, I plotted the data in a Perrin plot based on a similar linear
regression algorithm from assignment 3.

from matplotlib.pyplot import plot,title,show

from numpy import arange,array,ones,sin,sqrt,pi
from scipy import stats
from scipy.constants import Boltzmann

T = 293.15 #K
tau = 6.0#ns
r = [0.206, 0.217, 0.226, 0.249, 0.271, 0.292]
n = [1, 1.2, 1.41, 2.05, 3.62, 9.71]
x = [(T/i) for i in n]
y = [(1/i) for i in r]

slope, intercept, r_value, p_value, std_err = stats.linregress(x,y)

r0 = (1/intercept)
Vh = (tau*Boltzmann*intercept)/slope
Rho =Vh/(Boltzmann*T)
print 'r0 =', r0
print 'Vh =', Vh
print 'Rho =', Rho

y2=[(intercept, i) for i in y]
x2 = [(0.01,i) for i in x]
line = [slope*i+intercept for i in x]
plot(x2,y2,'o')
plot(x, line, 'r')
title('Perrin plot - Linear Fit for fluorescence polarization
anisotropy')
show()

The results returned were:

slope 0.00552375253977
intercept 3.24951609197
r0 = 0.307738128293
Vh 4.87325188896e-29
Rho 1.20405362512e-08

Thus for the protein in water we find:

Maximum Anisotropy: 0 = 0.308,
Hydrated Volume: = 4.87 1029 3,
and Rotational Correlation Time: = 12 .
These values seem reasonable based on similar values discussed in lecture and tutorials.

The linear regression in the plot in figure 1 is theoretically represented by the equation:
1 1
= (1 + )
0
PHY332 Assignment #5 Tim Henley 1000234506

Using the values from the linear regression the maximum anisotropy was found by:
1
0 =

The hydrated volume was found by:

=
0

And the rotational correlation time (RCT) was found by:

From the estimate in Lecture 11 (of 2.4 kDa per ns of RCT), the protein in question should have
a molecular weight in the neighbourhood of 29 kDa, which also seems reasonable.

The data points and regression were also used to make a rudimentary Perrin plot.

Figure 1. Perrin plot, 1/ vs. /, from linear regression based on the steady-state fluorescence
polarization anisotropy () of a dye labelled protein in water at 20 C with glycerol added to increase the
viscosity ().