FRET! April 27th

Reading

Lehninger, Box 12-3 (FRET special section), pages 448-449.

FRET (quick intro)

Campbell, Biophysical Techniques, pages 193-196.

From Lakowicz's Principles of Fluorerscence Spectroscopy

Wallrabe and Periasamy.
Imaging protein molecules using FRET and FLIM microscopy.
Curr. Opin. Biotech. 2005, 16, 19–27.

Homework

Cambell Problems: 5.5.1, 5.5.2, 5.5.3, 5.5.5, 5.5.6 (solutions)

 

Additional (Due At Exam Time): 15 pts

Access the article “Real-time monitoring of cyclic nucleotide signaling in neurons using genetically encoded FRET probes.” Brain Cell Biology. 2008, 36, 3.

Read the introduction, then look at the indicated figures to answer the questions below.

  1. The system you will be examining in the next question is shown in Figure 1B. What enzyme activity is being monitored? Is this upstream or downstream of cAMP signaling?
  2. Will FRET transfer increase or decrease during cAMP signaling?
  3. Now examine Figure 4. What ratio is being monitored? Is it higher or lower during cAMP signaling?
  4. How do the various neurotransmitters affect this ratio? What does this mean for cAMP signaling?
  5. Do all of the cells react the same? (How awesome is it that they can monitor all of them at once?!)

PreAssignment: FRET PreAssignment

In Class

In Class Worksheet ----- (solutions)

Slides