See the class slides the key point is that in

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See the class slides.  The key point is that in fluorescence, the excitation wavelength is  different from the emission wavelength, so there are no background photons at the  fluorescence wavelength. 4)  Tryptophan is a good fluorophore. 5)  (a) Three coenzymes that are good fluorophores: NADH, FAD, pyridoxyl phosphate (PLP). (b) NADH is a hydride donor, used in gluconeogensis (and many other pathways). FAD is an electron carrier found within the pyruvate dehydrogenase complex (PDC), and many  other places. PLP is found in the active site of transaminase enzymes (and many other places).
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6) (a)  Fluorescence quenching is the observation that the presence of some molecules  (fluorescence quenchers) can reduce the probability of fluorescence occuring, by providing an  alternate (non-radiative) pathway for relaxation to the electronic ground state. (b)  Tryptophan fluorescence versus (iodide) quencher concentration, for a protein that contains a  single  tryptophan that is on the protein’s surface : The plot would most likely have a negative slope (such as shown below). (c)  For tryptophan fluorescence versus (iodide) quencher concentration, for a protein that  contains a single tryptophan that is buried in the protein’s hydrophobic core: The plot would have a near-zero or slightly negative slope. For example, something like this:
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  • Spring '16
  • 100 mm, PLP, Hoff, 50 degrees, 5 mm

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