TY - JOUR
T1 - The monomeric G-proteins Rac1 and/or Cdc42 are required for the inhibition of voltage-dependent calcium current by bradykinin
AU - Wilk-Blaszczak, Malgorzata A.
AU - Singer, William D.
AU - Quill, Timothy
AU - Miller, Billy
AU - Frost, Jeffrey A.
AU - Sternweis, Paul C.
AU - Belardetti, Francesco
PY - 1997
Y1 - 1997
N2 - Although regulation of voltage-dependent calcium current (I(Ca,V)) by neurotransmitters is a ubiquitous mechanism among nerve cells, the signaling pathways involved are not well understood. We have determined previously that in a neuroblastomaglioma hybrid cell line (NG108-15), the heterotrimeric G- protein G13 mediates the inhibition of I(Ca,V) produced by bradykinin (BK) via an unknown mechanism. Various reports indicate that G13 can couple to RhoA, Rac1, and Cdc42, which are closely related members of the Rho family of monomeric G-proteins. We have investigated their role as signaling intermediates in the pathway used by BK to inhibit I(Ca,V). Using immunoblot analysis and the PCR, we found evidence that RhoA, Rac1, and Cdc42 all are expressed in NG108-15 cells. Intracellularly perfused recombinant Rho-GDI (an inhibitor of guanine nucleotide ex change specific for the Rho family) attenuated the inhibition of I(Ca,V) by BK. These findings indicate that activation of RhoA, Rac1, or Cdc42 may be required for the response to BK. To determine whether any of these monomeric G-proteins mediate the response to BK, we have intracellularly applied blocking antibodies specific for each of the candidate proteins. Only the anti-Rac1 antibody blocked the response to BK. In parallel experiments, peptides corresponding to the C-terminal regions of Rac1 and Cdc42 blocked the same response. These data indicate a novel functional contribution of Rac1 and possibly also of Cdc42 to the inhibition of I(Ca,V) by neurotransmitters.
AB - Although regulation of voltage-dependent calcium current (I(Ca,V)) by neurotransmitters is a ubiquitous mechanism among nerve cells, the signaling pathways involved are not well understood. We have determined previously that in a neuroblastomaglioma hybrid cell line (NG108-15), the heterotrimeric G- protein G13 mediates the inhibition of I(Ca,V) produced by bradykinin (BK) via an unknown mechanism. Various reports indicate that G13 can couple to RhoA, Rac1, and Cdc42, which are closely related members of the Rho family of monomeric G-proteins. We have investigated their role as signaling intermediates in the pathway used by BK to inhibit I(Ca,V). Using immunoblot analysis and the PCR, we found evidence that RhoA, Rac1, and Cdc42 all are expressed in NG108-15 cells. Intracellularly perfused recombinant Rho-GDI (an inhibitor of guanine nucleotide ex change specific for the Rho family) attenuated the inhibition of I(Ca,V) by BK. These findings indicate that activation of RhoA, Rac1, or Cdc42 may be required for the response to BK. To determine whether any of these monomeric G-proteins mediate the response to BK, we have intracellularly applied blocking antibodies specific for each of the candidate proteins. Only the anti-Rac1 antibody blocked the response to BK. In parallel experiments, peptides corresponding to the C-terminal regions of Rac1 and Cdc42 blocked the same response. These data indicate a novel functional contribution of Rac1 and possibly also of Cdc42 to the inhibition of I(Ca,V) by neurotransmitters.
KW - Cdc42
KW - G
KW - G-proteins
KW - NG108-15
KW - Rac1
KW - calcium channels
KW - modulation
KW - neuroblastoma- glioma
KW - neuropeptides
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UR - http://www.scopus.com/inward/citedby.url?scp=0030961897&partnerID=8YFLogxK
U2 - 10.1523/jneurosci.17-11-04094.1997
DO - 10.1523/jneurosci.17-11-04094.1997
M3 - Article
C2 - 9151726
AN - SCOPUS:0030961897
SN - 0270-6474
VL - 17
SP - 4094
EP - 4100
JO - Journal of Neuroscience
JF - Journal of Neuroscience
IS - 11
ER -