6533b82ffe1ef96bd12948c4
RESEARCH PRODUCT
8-Azido-adenosine 5'-triphosphate as a Photoaffinity Label for Bacterial F1 ATPase
Hans-jochen SchäferKlaus DosePeter Scheurichsubject
Macromolecular SubstancesUltraviolet RaysATPaseAffinity labelCooperativityBiochemistryMicrococcuschemistry.chemical_compoundAdenosine TriphosphateAdenine nucleotideUltraviolet lightMagnesiumNucleotideEdetic AcidAdenosine Triphosphataseschemistry.chemical_classificationPhotolysisbiologyAdenine NucleotidesChemistryAffinity LabelsBiochemistrybiology.proteinCalciumAdenosine triphosphateATP synthase alpha/beta subunitsdescription
1. 8-Azido-adenosine 5'-triphosphate (n83ATP) is a suitable photoaffinity label for F1 ATPase from Micrococcus luteus. The nucleotide is a substrate in the presence of bivalent cations and inhibits the enzyme irreversibly upon irradiation with ultraviolet light above 300 nm. 2. More than 80% of the label is covalently bound to the beta subunits in the presence of bivalent cations. Labeling and inactivation is decreased by protection with ADP, ATP or adenyl-5'-yl imidodiphosphate. To a much smaller degree the alpha subunits also become labeled. 3. n83AMP does not specifically bind to the beta subunits upon irradiation. Like n83ATP and n83ADP, it also labels the alpha subunits to a small extent. 4. The F1 ATPase is inactivated after a single beta subunit per F1 complex has become labeled. A cooperativity of the beta subunits carrying nucleotide binding sites is suggested.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1978-07-01 | European Journal of Biochemistry |