PROBLEMS
14.1 Write Beer’s law and define all of the terms.
14.2 What is the frequency of light with a wavelength of (a) 1 nm, (b) 500 nm, and (c) 3 cm?
Give the spectral classification of the radiation of each frequency.
14.3 What is the wavelength of light with a frequency of (a) 10
18 Hz, (b) 10
6 Hz, and (c) 10
12 Hz.
Give the spectral classification of the radiation of each wavelength.
14.4 A 0.1% solution of lysozyme gives an absorption of 2.7 at 280 nm. (a) If the concentration
is 0.01%, what absorption would be measured? (b) If an absorbance of 1.0 is measured,
what is the concentration?
14.5 Adenosine 5′-phosphate has an extinction coefficient of 1.54 × 10
4 M
−1 cm
−1 at 260 nm.
(a) If the concentration of the sample is 1 μM, what absorbance would be measured in a
1-cm cuvette? (b) If an absorbance of 0.1 is measured, what is the concentration?
14.6 If the extinction coefficient of a molecule is 1000 M
−1 cm
−1 and you measure an absorption
of 0.1 in a 1-cm cuvette, what is the concentration of the molecule?
CHAPTER 14
OPTICAL SPECTROSCOPY
315
and then cleaving the site. By labeling different ends of the DNA it is possible to get profiles for the distances between the two ends as a function
of parameters such as the enzyme concentration, and to derive possible
models for the arrangement of the enzyme on the DNA strands.
REFERENCES
Berg, J.M., Tymoczko, J.L., and Styer, L. (2007)
Biochemistry, 6th edn. W.H. Freeman, San
Francisco.
Brixner, T., Stenger, J., Vaswani, H.M. et al. (2005)
Two-dimensional spectroscopy of electronic couplings in photosynthesis. Nature 434, 625–8.
Churchill, M.E.A. (2003) Watching flipping junctions.
Nature Structural Biology 10, 73–5.
Katiliene, Z., Katilius, E., and Woodbury, N.W.
(2003) Single molecule detection of DNA looping
by NgoMIV restriction endonuclease. Biophysical
Journal 84, 4053–61.
Li, Y.F., Zhou, W., Blankenship, R.E., and Allen, J.P.
(1997) Crystal structure of the bacteriochlorophyll a protein from Chlorobium tepidum. Journal of
Molecular Biology 271, 456–71.
Matthews, B.W. and Fenna, R.E. (1980) Structure
of a green bacteriochlorophyll protein. Accounts of
Chemical Research 13, 309–17.
McKinney, S.A., Declais, A.C., Lilley, D.M.J., and
Ha, T. (2003) Structural dynamics of individual
Holliday junctions. Nature Structural Biology 10,
93–7.
Mollova, E.T. (2003) Single-molecule fluorescence
of nucleic acids. Current Opinion in Chemical Biology
6, 823–8.
Nie, S. and Zare, R.N. (1997) Optical detection of
single molecules. Annual Re9iew of Biophysics and
Biomolecular Structure 26, 567–96.
Olson, J.M. (1998) Chlorophyll organization and
function in green photosynthetic bacteria. Photochemistry and Photobiology 67, 61–75.
Peterman, E.J.G., Sosa, H., and Moerner, W.E.
(2004) Single-molecule fluorescence spectroscopy
and microscopy of biomolecular motors. Annual
Re9iew of Physical Chemistry 55, 79–96.
Weiss, S. (1999) Fluorescence spectroscopy of single
biomolecules. Science 283, 1676–83.
9781405124362_4_014.qxd 4/29/08 13:09 Page 315
14.1 Write Beer’s law and define all of the terms.
14.2 What is the frequency of light with a wavelength of (a) 1 nm, (b) 500 nm, and (c) 3 cm?
Give the spectral classification of the radiation of each frequency.
14.3 What is the wavelength of light with a frequency of (a) 10
18 Hz, (b) 10
6 Hz, and (c) 10
12 Hz.
Give the spectral classification of the radiation of each wavelength.
14.4 A 0.1% solution of lysozyme gives an absorption of 2.7 at 280 nm. (a) If the concentration
is 0.01%, what absorption would be measured? (b) If an absorbance of 1.0 is measured,
what is the concentration?
14.5 Adenosine 5′-phosphate has an extinction coefficient of 1.54 × 10
4 M
−1 cm
−1 at 260 nm.
(a) If the concentration of the sample is 1 μM, what absorbance would be measured in a
1-cm cuvette? (b) If an absorbance of 0.1 is measured, what is the concentration?
14.6 If the extinction coefficient of a molecule is 1000 M
−1 cm
−1 and you measure an absorption
of 0.1 in a 1-cm cuvette, what is the concentration of the molecule?
CHAPTER 14
OPTICAL SPECTROSCOPY
315
and then cleaving the site. By labeling different ends of the DNA it is possible to get profiles for the distances between the two ends as a function
of parameters such as the enzyme concentration, and to derive possible
models for the arrangement of the enzyme on the DNA strands.
REFERENCES
Berg, J.M., Tymoczko, J.L., and Styer, L. (2007)
Biochemistry, 6th edn. W.H. Freeman, San
Francisco.
Brixner, T., Stenger, J., Vaswani, H.M. et al. (2005)
Two-dimensional spectroscopy of electronic couplings in photosynthesis. Nature 434, 625–8.
Churchill, M.E.A. (2003) Watching flipping junctions.
Nature Structural Biology 10, 73–5.
Katiliene, Z., Katilius, E., and Woodbury, N.W.
(2003) Single molecule detection of DNA looping
by NgoMIV restriction endonuclease. Biophysical
Journal 84, 4053–61.
Li, Y.F., Zhou, W., Blankenship, R.E., and Allen, J.P.
(1997) Crystal structure of the bacteriochlorophyll a protein from Chlorobium tepidum. Journal of
Molecular Biology 271, 456–71.
Matthews, B.W. and Fenna, R.E. (1980) Structure
of a green bacteriochlorophyll protein. Accounts of
Chemical Research 13, 309–17.
McKinney, S.A., Declais, A.C., Lilley, D.M.J., and
Ha, T. (2003) Structural dynamics of individual
Holliday junctions. Nature Structural Biology 10,
93–7.
Mollova, E.T. (2003) Single-molecule fluorescence
of nucleic acids. Current Opinion in Chemical Biology
6, 823–8.
Nie, S. and Zare, R.N. (1997) Optical detection of
single molecules. Annual Re9iew of Biophysics and
Biomolecular Structure 26, 567–96.
Olson, J.M. (1998) Chlorophyll organization and
function in green photosynthetic bacteria. Photochemistry and Photobiology 67, 61–75.
Peterman, E.J.G., Sosa, H., and Moerner, W.E.
(2004) Single-molecule fluorescence spectroscopy
and microscopy of biomolecular motors. Annual
Re9iew of Physical Chemistry 55, 79–96.
Weiss, S. (1999) Fluorescence spectroscopy of single
biomolecules. Science 283, 1676–83.
9781405124362_4_014.qxd 4/29/08 13:09 Page 315