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Receptor specific for certain nucleotides stimulates inositol phosphate metabolism and Ca 2+ fluxes in A431 cells
Author(s) -
Gonzalez Fernando A.,
Alfonzo Ramona G.,
Toro Jorge R.,
Heppel Leon A.
Publication year - 1989
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041410320
Subject(s) - extracellular , inositol , nucleotide , biochemistry , inositol trisphosphate , inositol phosphate , cytosol , adenosine , biology , adenosine triphosphate , inositol trisphosphate receptor , chemistry , receptor , enzyme , gene
We have recently reported that extracellular ATP induces a transient rise in cytosolic free Ca 2+ ([Ca 2+ ] i ) in individual human epidermoid carcinoma A431 cells (Gonzalez et al: Journal of Cellular Physiology 135:269–276, 1988). We have now studied nucleotide specificity and desensitization for several early responses. Extracellular ATP (5–100 μM) caused the rapid formation of inositol trisphosphate and later its metabolites, inositol bisphosphate and inositol monophosphate. ATP also induced the efflux of 45 Ca 2+ from pre‐loaded cells. In addition, an increase in the rate of influx of 45 Ca 2+ stimulated by extracellular ATP was detected. Based on measurements of 45 Ca 2+ efflux and influx, desensitization studies, and chlortetracycline fluorimetry, we conclude that ATP mobilizes Ca 2+ from internal stores and also stimulates entry across the plasma membrane. These effects were also displayed by UTP and to a lesser extent by ITP, while other nucleoside triphosphates as well as ADP, AMP, and adenosine, were inactive. Furthermore, desensitization of the response to ATP and UTP was seen after prolonged exposure to either nucleotide. This was specific for the nucleotide receptor since a response to bradykinin was not affected by the ATP pretreatment, although pretreatment with phorbol ester inhibited responses to both the nucleotides and bradykinin. Quantitative data on rate of recovery from the desensitized state and the response of desensitized cells to greatly elevated levels of ATP are presented. Extracellular ATP stimulated another early change previously reported for epidermal growth factor, namely, the phosphorylation of an 81‐kDa cytoskeletal protein. The stimulation of these events involves an ATP receptor whose properties differ from other ATP receptors that have been described.