6533b7d7fe1ef96bd1267c5c
RESEARCH PRODUCT
8-N(3)-3'-biotinyl-ATP, a novel monofunctional reagent: differences in the F(1)- and V(1)-ATPases by means of the ATP analogue.
Hans-jochen SchäferHelmut WieczorekYasuo KagawaGerhard GrüberOlaf EgerJasminka Godovac-zimmermannÜNal Coskunsubject
Models MolecularVacuolar Proton-Translocating ATPasesTime FactorsUltraviolet RaysProtein subunitATPaseBiophysicsCoated vesicleBiotinPhotoaffinity LabelsPhotoaffinity LabelsBiochemistryCatalysischemistry.chemical_compoundAdenosine TriphosphateBiotinBacterial ProteinsManducaAnimalsBinding siteMolecular BiologyBinding SitesPhotoaffinity labelingbiologyChemistryCell BiologyProton-Translocating ATPasesBiochemistryModels ChemicalSpectrophotometrySpectrometry Mass Matrix-Assisted Laser Desorption-Ionizationbiology.proteinCattleGamma subunitProtein Bindingdescription
A novel photoaffinity label, 8-N(3)-3'-biotinyl-ATP, has been synthesized. The introduction of an additional biotin residue is advantageous for easy detection of labeled proteins. This could be first tested by reaction with the F(1)-ATPase from the thermophilic bacterium PS3 (TF(1)). UV irradiation of TF(1) in the presence of 8-N(3)-3'-biotinyl-ATP results in a nucleotide-dependent binding of the analogue in the noncatalytic alpha and the catalytic beta subunits of TF(1), demonstrating the suitability of this analogue as a potential photoaffinity label. Reaction with the V(1)-ATPase, however, led to labeling of subunit E, which has been suggested as a structural and functional homologue of the gamma subunit of the F-ATPases. MALDI-TOF mass spectrometry has been used to map the regions of subunit E involved in the binding of 8-N(3)-3'-biotinyl-ATP.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2001-09-01 | Biochemical and biophysical research communications |