title: Phosphorylation of the Ca2+ pump intermediate in intact red cells, isolated membranes and inside-out vesicles.
abstract: Ca2+-entry into intact red cells containing [32P]-ATP increases the phosphorylation of the 150 000 dalton polypeptide of the membrane. This phosphorylation occurs even in Mg2+-depleted red cells. Extracellular lanthanum applied during ATP-depletion further increases the Ca2+-induced phosphorylation. In fragmented membranes or resealed insideout vesicles (IOVs) membrane bound Mg2+ is sufficient to catalyze the phosphorylation of spectrin 2 and Band 3 polypeptides with low concentrations (less than micron of [32P]-ATP. In Ca-EDTA buffers one single polypeptide is phosphorylated which is located in the 150 000 molecular weight region. KmCa for phosphorylation is much lower (0.2 micron) than for active Ca2+ transport (40 micron) in IOVs. Lanthanum induced phosphorylation (up to 250 micron Lafree) is significantly greater than Ca2+-induced phosphorylation. Hg2+ inhibits both Ca2+ and La3+ induced phosphorylation. Ca2+-induced labelling can be rapidly "chased" by unlabelled ATP+Mg2+, but not with EGTA+Mg2+. Dephosphorylation in Ca2+ phosphorylated membranes and IOVs is significantly inhibited by La3+. It can be concluded that the mechanism of La3+ and Hg2+ inhibition of the Ca2+ pump is different in intact cells and isolated membranes or Iovs.
mesh_terms: D001693:Biological Transport, Active / Q000187:drug effects; D002118:Calcium / Q000097:blood* / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007811:Lanthanum / Q000494:pharmacology; D008565:Membrane Proteins / Q000097:blood*; D008628:Mercury / Q000494:pharmacology; D010750:Phosphoproteins / Q000097:blood*; D010766:Phosphorylation
chemical_list: D008565:Membrane Proteins; D010750:Phosphoproteins; D007811:Lanthanum; D008628:Mercury; D002118:Calcium