PMID- 22117552
OWN - NLM
STAT- MEDLINE
DCOM- 20120315
LR  - 20131121
IS  - 1608-3040 (Electronic)
IS  - 0006-2979 (Linking)
VI  - 76
IP  - 11
DP  - 2011 Nov
TI  - Characteristics of protection by MgADP and MgATP of alpha3beta3gamma subcomplex of 
      thermophilic Bacillus PS3 betaY341W-mutant F1-ATPase from inhibition by 
      7-chloro-4-nitrobenz-2-oxa-1,3-diazole support a bi-site mechanism of catalysis.
PG  - 1253-61
LID - 10.1134/S0006297911110071 [doi]
AB  - MgADP and MgATP binding to catalytic sites of betaY341W-alpha(3)beta(3)gamma subcomplex of 
      F(1)-ATPase from thermophilic Bacillus PS3 has been assessed using their effect 
      on the enzyme inhibition by 7-chloro-4-nitrobenz-2-oxa-1,3-diazole (NBD-Cl). It 
      was assumed that NBD-Cl can inhibit only when catalytic sites are empty, and 
      inhibition is prevented if a catalytic site is occupied with a nucleotide. In the 
      absence of an activator, MgADP and MgATP protect betaY341W-alpha(3)beta(3)gamma subcomplex from 
      inhibition by NBD-Cl by binding to two catalytic sites with an affinity of 37 microM 
      and 12 mM, and 46 microM and 15 mM, respectively. In the presence of an activator 
      lauryldimethylamine-N-oxide (LDAO), MgADP protects betaY341W-alpha(3)beta(3)gamma subcomplex 
      from inhibition by NBD-Cl by binding to a catalytic site with a K(d) of 12 mM. 
      Nucleotide binding to a catalytic site with affinity in the millimolar range has 
      not been previously revealed in the fluorescence quenching experiments with 
      betaY341W-alpha(3)beta(3)gamma subcomplex. In the presence of activators LDAO or selenite, 
      MgATP protects betaY341W-alpha(3)beta(3)gamma subcomplex from inhibition by NBD-Cl only 
      partially, and the enzyme remains sensitive to inhibition by NBD-Cl even at MgATP 
      concentrations that are saturating for ATPase activity. The results support a 
      bi-site mechanism of catalysis by F(1)-ATPases.
FAU - Milgrom, Y M
AU  - Milgrom YM
AD  - Department of Biochemistry and Molecular Biology, State University of New York, 
      Upstate Medical University, Syracuse, New York 13210, USA. milgromy@upstate.edu
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Biochemistry (Mosc)
JT  - Biochemistry. Biokhimiia
JID - 0376536
RN  - 0 (7-chloro-4-nitrobenz-2-oxa-1,3-diazole)
RN  - 0 (Dimethylamines)
RN  - 0 (Mutant Proteins)
RN  - 0 (Nitrobenzenes)
RN  - 0 (Oxazoles)
RN  - 0 (Protein Subunits)
RN  - 4F6FC4MI8W (dodecyldimethylamine oxide)
RN  - 61D2G4IYVH (Adenosine Diphosphate)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 3.6.1.- (Bacterial Proton-Translocating ATPases)
SB  - IM
MH  - Adenosine Diphosphate/*chemistry
MH  - Adenosine Triphosphate/*chemistry
MH  - Bacillus/*enzymology
MH  - Bacterial Proton-Translocating ATPases/antagonists & inhibitors/*chemistry
MH  - Binding Sites
MH  - Catalysis
MH  - Catalytic Domain
MH  - Dimethylamines/chemistry
MH  - Kinetics
MH  - Mutant Proteins/*chemistry/metabolism
MH  - Nitrobenzenes/chemistry
MH  - Oxazoles/chemistry
MH  - Protein Subunits/chemistry
EDAT- 2011/11/29 06:00
MHDA- 2012/03/16 06:00
CRDT- 2011/11/29 06:00
PHST- 2011/11/29 06:00 [entrez]
PHST- 2011/11/29 06:00 [pubmed]
PHST- 2012/03/16 06:00 [medline]
AID - BCM76111556 [pii]
AID - 10.1134/S0006297911110071 [doi]
PST - ppublish
SO  - Biochemistry (Mosc). 2011 Nov;76(11):1253-61. doi: 10.1134/S0006297911110071.