PMID- 23414455
OWN - NLM
STAT- MEDLINE
DCOM- 20130903
LR  - 20211021
IS  - 1520-5126 (Electronic)
IS  - 0002-7863 (Print)
IS  - 0002-7863 (Linking)
VI  - 135
IP  - 9
DP  - 2013 Mar 6
TI  - Mapping the nucleotide binding site of uncoupling protein 1 using atomic force 
      microscopy.
PG  - 3640-6
LID - 10.1021/ja312550k [doi]
AB  - A tight regulation of proton transport in the inner mitochondrial membrane is 
      crucial for physiological processes such as ATP synthesis, heat production, or 
      regulation of the reactive oxygen species as proposed for the uncoupling protein 
      family members (UCP). Specific regulation of proton transport is thus becoming 
      increasingly important in the therapy of obesity and inflammatory, 
      neurodegenerative, and ischemic diseases. We and other research groups have shown 
      previously that UCP1- and UCP2-mediated proton transport is inhibited by purine 
      nucleotides. Several hypotheses have been proposed to explain the inhibitory 
      effect of ATP, although structural details are still lacking. Moreover, the 
      unresolved mystery is how UCP operates in vivo despite the permanent presence of 
      high (millimolar) concentrations of ATP in mitochondria. Here we use the 
      topographic and recognition (TREC) mode of an atomic force microscope to 
      visualize UCP1 reconstituted into lipid bilayers and to analyze the ATP-protein 
      interaction at a single molecule level. The comparison of recognition patterns 
      obtained with anti-UCP1 antibody and ATP led to the conclusion that the ATP 
      binding site can be accessed from both sides of the membrane. Using cantilever 
      tips with different cross-linker lengths, we determined the location of the 
      nucleotide binding site inside the membrane with 1 A precision. Together with the 
      recently published NMR structure of a UCP family member (Berardi et al. Nature, 
      2011, 476, 109-113), our data provide a valuable insight into the mechanism of 
      the nucleotide binding and pave the way for new pharmacological approaches 
      against the diseases mentioned above.
FAU - Zhu, Rong
AU  - Zhu R
AD  - Institute for Biophysics, Johannes Kepler University, Linz, Austria.
FAU - Rupprecht, Anne
AU  - Rupprecht A
FAU - Ebner, Andreas
AU  - Ebner A
FAU - Haselgrubler, Thomas
AU  - Haselgrubler T
FAU - Gruber, Hermann J
AU  - Gruber HJ
FAU - Hinterdorfer, Peter
AU  - Hinterdorfer P
FAU - Pohl, Elena E
AU  - Pohl EE
LA  - eng
GR  - F 3503/FWF_/Austrian Science Fund FWF/Austria
GR  - P 25357/FWF_/Austrian Science Fund FWF/Austria
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130226
PL  - United States
TA  - J Am Chem Soc
JT  - Journal of the American Chemical Society
JID - 7503056
RN  - 0 (Ion Channels)
RN  - 0 (Mitochondrial Proteins)
RN  - 0 (Purine Nucleotides)
RN  - 0 (Uncoupling Protein 1)
SB  - IM
MH  - Binding Sites
MH  - Ion Channels/antagonists & inhibitors/*chemistry
MH  - Microscopy, Atomic Force
MH  - Mitochondrial Proteins/antagonists & inhibitors/*chemistry
MH  - Models, Molecular
MH  - Purine Nucleotides/*chemistry/metabolism
MH  - Uncoupling Protein 1
PMC - PMC3593612
EDAT- 2013/02/19 06:00
MHDA- 2013/09/04 06:00
PMCR- 2013/03/11
CRDT- 2013/02/19 06:00
PHST- 2013/02/19 06:00 [entrez]
PHST- 2013/02/19 06:00 [pubmed]
PHST- 2013/09/04 06:00 [medline]
PHST- 2013/03/11 00:00 [pmc-release]
AID - 10.1021/ja312550k [doi]
PST - ppublish
SO  - J Am Chem Soc. 2013 Mar 6;135(9):3640-6. doi: 10.1021/ja312550k. Epub 2013 Feb 
      26.