Transient inhibition of capacitative calcium entry in human neutrophils by a monoclonal antibody directed against a 19-kDa antigen.
P1C3 is a monoclonal antibody that binds p19, a novel neutrophil activation antigen that translocates to the cell surface upon neutrophil activation. We find that P1C3 inhibits capacitative Ca2+ entry, induced by emptying the intracellular Ca2+ stores with thapsigargin. The effect is transient, reaching its maximum at 30-60 s, but becomes permanent upon pretreatment of the cells with the protein phosphatase inhibitor calyculin A, suggesting the involvement of protein phosphorylation. The inhibitory action is similar to the one reported previously for the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP), although the transduction mechanism may be different. Inhibition of Ca2+ entry by fMLP was prevented by pretreatment with pertussis toxin, whereas inhibition by P1C3 was not. Pretreatment with cholera toxin had no effect. This suggests that the effect of P1C3 may not be mediated by a heterotrimeric G protein. Tyrosine kinase inhibitors did not prevent inhibition by either fMLP or P1C3. Phospholipase C activation seems not to be involved as P1C3, contrarily to fMLP, was unable to induce Ca2+ release from the intracellular Ca2+ stores.
Alonso
M T
MT
Instituto de Biologia y Genetica Molecular, Universidad de Valladolid y Consejo Superior de Investigaciones Científicas, Spain.
Mollinedo
F
F
Barasoain
I
I
Alvarez
J
J
Garcia-Sancho
J
J
D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology*; D002118:Calcium / Q000097:blood* / Q000493:pharmacokinetics; D018922:HL-60 Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology; D008868:Microtubule Proteins; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D018375:Neutrophil Activation; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D010750:Phosphoproteins / Q000276:immunology* / Q000378:metabolism; D050777:Stathmin; D019284:Thapsigargin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008868:Microtubule Proteins; D010750:Phosphoproteins; C493995:STMN1 protein, human; D050777:Stathmin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019284:Thapsigargin; D002118:Calcium