8-N-3-3 `-biotnyl-ATP, a novel monofunctional reagent: Differences in the F-1- and V-1-ATPases by means of the ATP analogue
Autor(en): | Schafer, HJ Coskun, U Eger, O Godovac-Zimmermann, J Wieczorek, H Kagawa, Y Gruber, G |
Stichwörter: | 5'-TRIPHOSPHATE; 8-AZIDO-ATP; Biochemistry & Molecular Biology; Biophysics; COATED VESICLE; F-1-ATPase; GAMMA-SUBUNIT; H+-translocating vacuolar-type ATPase; HEART MITOCHONDRIAL ATPASE; Manduca sexta; NUCLEOTIDE-BINDING-SITES; PHOTOAFFINITY LABEL; photoaffinity labeling; RESOLUTION; thermophilic bacterium PS3; V-1-ATPase; V-ATPASE; VACUOLAR (H+)-ATPASE | Erscheinungsdatum: | 2001 | Herausgeber: | ACADEMIC PRESS INC ELSEVIER SCIENCE | Journal: | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | Volumen: | 286 | Ausgabe: | 5 | Startseite: | 1218 | Seitenende: | 1227 | Zusammenfassung: | 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 (TF1). UV irradiation of TF1 in the presence of 8-N-3-3'-biotinyl-ATP results in a nucleotide-dependent binding of the analogue in the noncatalytic a and the catalytic beta subunits of TF1, 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'-biotiinyl-ATP. (C) 2001 Academic Press. |
ISSN: | 0006291X | DOI: | 10.1006/bbrc.2001.5502 |
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geprüft am 04.05.2024