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Topic 4 (abridged)
Biophysics
Energy and Absorption
2 t 2 sin 2 E b E a w t
~
b a E0 2
Pb C b
2
2
2 E b
w t
Ea
2
2
• Absorption is
proportional to
~
a b E0
• Get resonance at hf
= ħw = Eb – Ea
www.andor.com/chemistry/?app=189
Absorption Broadening
•Every electronic state
(eg S0 and S1) has
several vibrational and
rotational substates.
•Each of the sharp peaks
is further broadened by
Doppler shifts and
solvent effects and
quantum probabilities.
pubs.rsc.org/ej/DT/2000/B002123G/
Extinction Coefficient
monochromator l I
I0
Sample with
Light Source pathlength l Detector
Or
l I1
I0
Beam Reference
monochromator Detector
Splitter
Light Source l I2
I0
Sample with
pathlength l Detector
Blood
http://life.nthu.edu.tw/~lslpc/biophys/hyperchorism.gif
Chemical Kinetics
Zero Order
1.40E+00
1.20E+00
1.00E+00 Reaction of nitrite
Absorption
C 8.00E-01
6.00E-01
A
B
with
4.00E-01 deoxyhemoglobin
2.00E-01
0.00E+00
450 500 550 600 650 700
Wavelength (nm)
t
k
A Z
• -dCA/dt = kCA , CA = CA0 e-kt
• t1/2 = ln(2)/k; t = 1/k = lifetime NO binding to Hb
Chemical Kinetics
Second Order
AB
Pr oducts k
Cooperative Binding of
Oxygen
Linked to quaternary
structure
Explained by MWC Model
On the Nature of Allosteric
Transitions:
A Plausible Model
Jacques Monod, Jeffries Wyman,
Jean-Pierre Changux
J. Mol. Biol. 1965
Time Resolved Optical Density
TROD
Stopped-flow Pump-probe
http://fig.cox.miami.edu/~cmallery/255/255enz/ecb3x28.jpg
www.math.tau.ac.il/~nin/Courses/BioInfo04/svd.ppt
http://upload.wikimedia.org/wikipedia/com
mons/thumb/d/d1/Circular_dichroism.png/6
22px-Circular_dichroism.png