PMID- 27104936
OWN - NLM
STAT- MEDLINE
DCOM- 20170628
LR  - 20170628
IS  - 1463-9084 (Electronic)
IS  - 1463-9076 (Linking)
VI  - 18
IP  - 18
DP  - 2016 May 14
TI  - A simulation-guided fluorescence correlation spectroscopy tool to investigate the 
      protonation dynamics of cytochrome c oxidase.
PG  - 12877-85
LID - 10.1039/c5cp07925j [doi]
AB  - Fluorescence correlation spectroscopy (FCS) is a single molecule based technique 
      to temporally resolve rate-dependent processes by correlating the fluorescence 
      fluctuations of individual molecules traversing through a confocal volume. In 
      addition, chemical processes like protonation or intersystem crossing can be 
      monitored in the sub-microsecond range. FCS thereby provides an excellent tool 
      for investigations of protonation dynamics in proton pumps like cytochrome c 
      oxidase (CcO). To achieve this, the pH-dependent fluorescent dye fluorescein was 
      attached as a protonation probe to the CcO surface via site-specific labeling of 
      single reactive cysteines that are located close to the entry point of a proton 
      input channel (K-pathway). The analysis of protonation dynamics is complicated by 
      overlapping triplet and protonation rates of the fluorophore. A Monte Carlo 
      simulation based algorithm was developed to facilitate discrimination of these 
      temporally overlapping processes thus allowing for improved protonation reaction 
      rate determination. Using this simulation-guided approach we determined precise 
      local proton association and dissociation rates and provide information about 
      protein surface effects, such as proton collecting antennae, on the transport 
      properties of proton transfer channels.
FAU - Wolf, A
AU  - Wolf A
AD  - Department of Physics, Freie Universitat Berlin, 14195 Berlin, Germany. 
      ulrike.alexiev@fu-berlin.de.
FAU - Schneider, C
AU  - Schneider C
FAU - Kim, T-Y
AU  - Kim TY
FAU - Kirchberg, K
AU  - Kirchberg K
FAU - Volz, P
AU  - Volz P
FAU - Alexiev, U
AU  - Alexiev U
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160422
PL  - England
TA  - Phys Chem Chem Phys
JT  - Physical chemistry chemical physics : PCCP
JID - 100888160
RN  - 0 (Protons)
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
SB  - IM
MH  - Electron Transport Complex IV/*chemistry
MH  - Fluorescence
MH  - Models, Molecular
MH  - Monte Carlo Method
MH  - Paracoccus denitrificans/chemistry/*enzymology
MH  - Protons
MH  - Spectrometry, Fluorescence/*methods
EDAT- 2016/04/23 06:00
MHDA- 2017/06/29 06:00
CRDT- 2016/04/23 06:00
PHST- 2016/04/23 06:00 [entrez]
PHST- 2016/04/23 06:00 [pubmed]
PHST- 2017/06/29 06:00 [medline]
AID - 10.1039/c5cp07925j [doi]
PST - ppublish
SO  - Phys Chem Chem Phys. 2016 May 14;18(18):12877-85. doi: 10.1039/c5cp07925j. Epub 
      2016 Apr 22.