TY - JOUR
T1 - Delineation of molecular determinants for FR900359 inhibition of Gq/11 unlocks inhibition of Gas
AU - Boesgaard, Michael W.
AU - Harpsøe, Kasper
AU - Malmberg, Michelle
AU - Underwood, Christina R.
AU - Inoue, Asuka
AU - Mathiesen, Jesper M.
AU - König, Gabriele M.
AU - Kostenis, Evi
AU - Gloriam, David E.
AU - Bräuner-Osborne, Hans
N1 - Funding Information:
Funding and additional information—This work was supported by Independent Research Fund Denmark | Medical Sciences 8020-00280B (to D. E. G.); Lundbeck Foundation Grants R163-2013-16327 (to D. E. G.), R178-2014-2060 (to H. B.-O. and D. E. G.), and R171-2014-505 (to C. R. U); Novo Nordisk Foundation Grant NNF18OC0031226; German Research Foundation (DFG) Grants KO 1582/10-1 and KO 1582/10-2 within FOR2372 (to E. K.) as well as KO 902/17-1 and KO 902/17-2 (to G. M. K.); Japan Society for the Promotion of Science (JSPS) KAKENHI Grant 17K08264 (to A. I.); and Japan Agency for Medical Research and Development (AMED) Grants PRIME JP17gm5910013 and LEAP JP17gm0010004 (to A. I.).
PY - 2020/10/2
Y1 - 2020/10/2
N2 - Heterotrimeric G proteins are essential mediators of intracellular signaling of G protein-coupled receptors. The Gq/11 subfamily consists of Gq, G11, G14, and G16 proteins, of which all but G16 are inhibited by the structurally related natural products YM-254890 and FR900359. These inhibitors act by preventing the GDP/GTP exchange, which is necessary for activation of all G proteins. A homologous putative binding site for YM-254890/ FR900359 can also be found in members of the other three G protein families, Gs, Gi/o, and G12/13, but none of the published analogs of YM-254890/FR900359 have shown any inhibitory activity for any of these. To explain why the YM-254890/ FR900359 scaffold only inhibits Gq/11/14, the present study delineated the molecular selectivity determinants by exchanging amino acid residues in the YM-254890/FR900359-binding site in Gq and Gs. We found that the activity of a Gs mutant with a Gq-like binding site for YM-254890/FR900359 can be inhibited by FR900359, and a minimum of three mutations are necessary to introduce inhibition in Gs. In all, this suggests that although the YM-254890/FR900359 scaffold has proven unsuccessful to derive Gs, Gi/o, and G12/13 inhibitors, the mechanism of inhibition between families of G proteins is conserved, opening up the possibility of targeting by other, novel inhibitor scaffolds. In lack of a selective Gas inhibitor, FR900359-sensitive Gas mutants may prove useful in studies where delicate control over Gas signaling would be of the essence.
AB - Heterotrimeric G proteins are essential mediators of intracellular signaling of G protein-coupled receptors. The Gq/11 subfamily consists of Gq, G11, G14, and G16 proteins, of which all but G16 are inhibited by the structurally related natural products YM-254890 and FR900359. These inhibitors act by preventing the GDP/GTP exchange, which is necessary for activation of all G proteins. A homologous putative binding site for YM-254890/ FR900359 can also be found in members of the other three G protein families, Gs, Gi/o, and G12/13, but none of the published analogs of YM-254890/FR900359 have shown any inhibitory activity for any of these. To explain why the YM-254890/ FR900359 scaffold only inhibits Gq/11/14, the present study delineated the molecular selectivity determinants by exchanging amino acid residues in the YM-254890/FR900359-binding site in Gq and Gs. We found that the activity of a Gs mutant with a Gq-like binding site for YM-254890/FR900359 can be inhibited by FR900359, and a minimum of three mutations are necessary to introduce inhibition in Gs. In all, this suggests that although the YM-254890/FR900359 scaffold has proven unsuccessful to derive Gs, Gi/o, and G12/13 inhibitors, the mechanism of inhibition between families of G proteins is conserved, opening up the possibility of targeting by other, novel inhibitor scaffolds. In lack of a selective Gas inhibitor, FR900359-sensitive Gas mutants may prove useful in studies where delicate control over Gas signaling would be of the essence.
UR - http://www.scopus.com/inward/record.url?scp=85092682336&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85092682336&partnerID=8YFLogxK
U2 - 10.1074/jbc.RA120.013002
DO - 10.1074/jbc.RA120.013002
M3 - Article
C2 - 32753482
AN - SCOPUS:85092682336
VL - 295
SP - 13850
EP - 13861
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 40
ER -