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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