PMID- 32977390
OWN - NLM
STAT- MEDLINE
DCOM- 20210223
LR  - 20210223
IS  - 1422-0067 (Electronic)
IS  - 1422-0067 (Linking)
VI  - 21
IP  - 19
DP  - 2020 Sep 23
TI  - Lipid Composition Affects the Efficiency in the Functional Reconstitution of the 
      Cytochrome c Oxidase.
LID - 10.3390/ijms21196981 [doi]
LID - 6981
AB  - The transmembrane protein cytochrome c oxidase (CcO) is the terminal oxidase in 
      the respiratory chain of many aerobic organisms and catalyzes the reduction of 
      dioxygen to water. This process maintains an electrochemical proton gradient 
      across the membrane hosting the oxidase. CcO is a well-established model enzyme 
      in bioenergetics to study the proton-coupled electron transfer reactions and 
      protonation dynamics involved in these processes. Its catalytic mechanism is 
      subject to ongoing intense research. Previous research, however, was mainly 
      focused on the turnover of oxygen and electrons in CcO, while studies reporting 
      proton turnover rates of CcO, that is the rate of proton uptake by the enzyme, 
      are scarce. Here, we reconstitute CcO from R. sphaeroides into liposomes 
      containing a pH sensitive dye and probe changes of the pH value inside single 
      proteoliposomes using fluorescence microscopy. CcO proton turnover rates are 
      quantified at the single-enzyme level. In addition, we recorded the distribution 
      of the number of functionally reconstituted CcOs across the proteoliposome 
      population. Studies are performed using proteoliposomes made of native lipid 
      sources, such as a crude extract of soybean lipids and the polar lipid extract of 
      E. coli, as well as purified lipid fractions, such as phosphatidylcholine 
      extracted from soybean lipids. It is shown that these lipid compositions have 
      only minor effects on the CcO proton turnover rate, but can have a strong impact 
      on the reconstitution efficiency of functionally active CcOs. In particular, our 
      experiments indicate that efficient functional reconstitution of CcO is strongly 
      promoted by the addition of anionic lipids like phosphatidylglycerol and 
      cardiolipin.
FAU - Hugentobler, Katharina Gloria
AU  - Hugentobler KG
AUID- ORCID: 0000-0002-3551-3135
AD  - Institute of Chemistry and Biochemistry, Emmy-Noether Group "Bionanointerfaces", 
      Freie Universitat Berlin, Arnimallee 22, 14195 Berlin, Germany.
FAU - Heinrich, Dorothea
AU  - Heinrich D
AD  - Department of Physics, Genetic Biophysics, Freie Universitat Berlin, Arnimallee 
      22, 14195 Berlin, Germany.
FAU - Berg, Johan
AU  - Berg J
AD  - Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural 
      Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.
FAU - Heberle, Joachim
AU  - Heberle J
AUID- ORCID: 0000-0001-6321-2615
AD  - Department of Physics, Experimental Molecular Biophysics, Freie Universitat 
      Berlin, Arnimallee 22, 14195 Berlin, Germany.
FAU - Brzezinski, Peter
AU  - Brzezinski P
AD  - Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural 
      Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.
FAU - Schlesinger, Ramona
AU  - Schlesinger R
AUID- ORCID: 0000-0002-7716-4439
AD  - Department of Physics, Genetic Biophysics, Freie Universitat Berlin, Arnimallee 
      22, 14195 Berlin, Germany.
FAU - Block, Stephan
AU  - Block S
AUID- ORCID: 0000-0002-2947-0837
AD  - Institute of Chemistry and Biochemistry, Emmy-Noether Group "Bionanointerfaces", 
      Freie Universitat Berlin, Arnimallee 22, 14195 Berlin, Germany.
LA  - eng
GR  - BL1514-1/1/Deutsche Forschungsgemeinschaft/
GR  - SFB 1078 (A1, A6, B4)/Deutsche Forschungsgemeinschaft/
PT  - Journal Article
DEP - 20200923
PL  - Switzerland
TA  - Int J Mol Sci
JT  - International journal of molecular sciences
JID - 101092791
RN  - 0 (Bacterial Proteins)
RN  - 0 (Liposomes)
RN  - 0 (Membrane Lipids)
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
SB  - IM
MH  - Bacterial Proteins/*chemistry
MH  - Electron Transport Complex IV/*chemistry
MH  - Hydrogen-Ion Concentration
MH  - Liposomes
MH  - Membrane Lipids/*chemistry
MH  - Rhodobacter sphaeroides/*enzymology
PMC - PMC7583929
OTO - NOTNLM
OT  - electron transfer
OT  - membrane protein
OT  - proton pump
OT  - proton translocation
OT  - single enzyme fluorescence microscopy
OT  - single molecule
COIS- The authors declare no conflict of interest.
EDAT- 2020/09/27 06:00
MHDA- 2021/02/24 06:00
PMCR- 2020/10/01
CRDT- 2020/09/26 01:00
PHST- 2020/08/24 00:00 [received]
PHST- 2020/09/15 00:00 [revised]
PHST- 2020/09/18 00:00 [accepted]
PHST- 2020/09/26 01:00 [entrez]
PHST- 2020/09/27 06:00 [pubmed]
PHST- 2021/02/24 06:00 [medline]
PHST- 2020/10/01 00:00 [pmc-release]
AID - ijms21196981 [pii]
AID - ijms-21-06981 [pii]
AID - 10.3390/ijms21196981 [doi]
PST - epublish
SO  - Int J Mol Sci. 2020 Sep 23;21(19):6981. doi: 10.3390/ijms21196981.