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J. Biol. Chem., Vol. 259, Issue 8, 4910-4916, 04, 1984
S Kassis, RC Henneberry and PH Fishman
HeLa cells, when exposed to 5 mM sodium butyrate, increased their
responsiveness to isoproterenol and their number of beta-receptors. As
untreated HeLa cells have a substantial number of receptors but respond
poorly to isoproterenol, the effect of butyrate could be due to
quantitative or qualitative changes in beta-receptors or other components
of the adenylate cyclase system. Receptors were analyzed by
membrane/membrane and membrane/cell fusion techniques. HeLa donor
membranes, treated to inactivate regulatory and catalytic components of
adenylate cyclase, were fused with Fc cells, which lack beta-receptors.
Isoproterenol-stimulated adenylate cyclase activity in the fusates was
proportional to the number of receptors present. There appeared to be only
quantitative but not qualitative differences in beta-receptors from control
and butyrate-treated HeLa. Prostaglandin E1 receptors from neuroblastoma
cell membranes were similarly coupled to HeLa adenylate cyclase. The hybrid
prostaglandin E1-stimulated activity was lower when acceptor membranes were
from control HeLa than when they were from butyrate-treated HeLa cells.
These results suggested that butyrate was altering the ability of the
regulatory component to interact with receptors. HeLa membranes were
extracted with sodium cholate and the extracts used to reconstitute
effector-stimulated adenylate cyclase activity in S49 cyc- membranes, which
lack a functional regulatory component. Whereas extracts from control and
butyrate-treated HeLa were equally effective in restoring NaF-stimulated
activity in cyc- membranes, extracts from control HeLa were less efficient
in reconstituting isoproterenol- and prostaglandin E1-stimulated
activities. We conclude that the poor response of control HeLa to beta-
agonists is due to a limited activity of the regulatory component but not
the receptor. Butyrate induces quantitative changes in the receptor and
qualitative changes in the regulatory component that facilitate its ability
to couple to receptors but do not alter its ability to interact with the
catalytic component of adenylate cyclase.
Induction of catecholamine-responsive adenylate cyclase in HeLa cells by sodium butyrate. Evidence for a more efficient stimulatory regulatory component
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