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Frster Resonance Energy Transfer principle

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The efficiency E of the resonance energy transfer from a fluorescent donor to an ... Sonya Kuznetsova, MetProt Grp 2005. Oct. 13, 2005. GW Canters, Leiden Univ. 3 ... – PowerPoint PPT presentation

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Title: Frster Resonance Energy Transfer principle


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Förster Resonance Energy Transfer principle
The efficiency E of the resonance energy transfer
from a fluorescent donor to an acceptor molecule
is E R06/(R6R06), R
donor-acceptor distance, R0 Förster radius
R06 ?2 J(?), ?2 orientation factor,
J(?) - spectral overlap between the donor
fluorescence and acceptor absorption
Sonya Kuznetsova, MetProt Grp 2005
3
R0 is constant and determined by the spectral
properties of the donor and the acceptor. The
donor-acceptor distance R is changing and can be
determined from the fluorescence intensity.
Sonya Kuznetsova, MetProt Grp 2005
4
3(2002)908
5
energy transfer
6
-
O
S
SO
-
3
3
42
N
N
12
O

N



N
O
O
27
7
ChemPhysChem (2005), 61381-1386.
J. Biol. Inorg.Chem (2005)
8
Single Molecules Wide Field Set-up
Ralf Schmauder, 2005
Stokes- shift
9
Single Molecules Raw Data
200
Ralf Schmauder, 2005
100
0
5 mm
10
Single Molecules brightness
ChemPhysChem (2005), 61381-1386.
oxidized
reduced
11
Control Zn-Azurin
ChemPhysChem (2005), 61381-1386.
12
Close up
ChemPhysChem (2005), 61381-1386.
hnA
hnE
1 fl 10-15 l
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The raw data chemistry
ChemPhysChem (2005), 61381-1386.
Detector
Laser
time
fluorescence
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Azurin Data Cu
ChemPhysChem (2005), 61381-1386.
15
Azurin Data Zn
ChemPhysChem (2005), 61381-1386.
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Cholesterol oxidase
Science 282(1998)1877
18
C.antarctica Lipase B
Angew. Chem. 44(2005)560
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