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Identification of transmembrane domains ... ctivatable phospholipidic probe
Identification of transmembrane domains of the red cell calcium pump with a new photoactivatable phospholipidic probe
5378
The membrane-associated regions of the human erythrocyte Ca2+ pump were investigated by hydrophobic photolabeling. Purified Ca2+ pump was reconstituted in asolectin vesicles loaded with [3H]DIPETPD, a photochemical probe designed to label deeply into the hydrophobic core of the lipid bilayer (Delfino et al. J. Am. Chem. Soc. 115, 3458-3474, 1993). After photolysis and SDS-PAGE analysis, a significant light-dependent labeling of the Ca2+ pump was found. Controlled proteolysis of the photoadduct with trypsin or protease V8 followed by SDS-PAGE and immunoblotting yielded individual labeled fragments. The labeling pattern indicated the existence of three sequential clusters of transmembrane regions, consistent with the current model for the topography of this enzyme.
Castello PR, Caride AJ, González Flecha FL, Fernández HN, Rossi JP, Delfino JM
Biochemical and biophysical research communications
1994-05-30 00:00
201
1
194-200
Calcium-Transporting ATPases,Erythrocytes,Humans,Membrane Glycoproteins,Phospholipids,Serine Endopeptidases,Structure-Activity Relationship,Trypsin,Membrane Glycoproteins,Phospholipids,Serine Endopeptidases,glutamyl endopeptidase,Trypsin,Calcium-Transporting ATPases
IQUIFIB, Instituto de Química y Fisicoquímica Biológicas (UBA-CONICET), Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina.
Biochem. Biophys. Res. Commun.
0006-291X
S0006291X84716883
0
False
8198574
