Massive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.

TRPM6 and TRPM7 are bifunctional proteins expressing a TRP channel fused to an atypical alpha-kinase domain. While the gating properties of TRPM6 and TRPM7 channels have been studied in detail, little is known about the mechanisms regulating kinase activity. Recently, we found that TRPM7 associates...

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Autores principales:	Kristopher Clark, Jeroen Middelbeek, Nick A Morrice, Carl G Figdor, Edwin Lasonder, Frank N van Leeuwen
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2008
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7698f9dc761844ae84244a5b619b0d9f
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spelling	oai:doaj.org-article:7698f9dc761844ae84244a5b619b0d9f2021-11-25T06:12:58ZMassive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.1932-620310.1371/journal.pone.0001876https://doaj.org/article/7698f9dc761844ae84244a5b619b0d9f2008-03-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/18365021/?tool=EBIhttps://doaj.org/toc/1932-6203TRPM6 and TRPM7 are bifunctional proteins expressing a TRP channel fused to an atypical alpha-kinase domain. While the gating properties of TRPM6 and TRPM7 channels have been studied in detail, little is known about the mechanisms regulating kinase activity. Recently, we found that TRPM7 associates with its substrate myosin II via a kinase-dependent mechanism suggesting a role for autophosphorylation in substrate recognition. Here, we demonstrate that the cytosolic C-terminus of TRPM7 undergoes massive autophosphorylation (32+/-4 mol/mol), which strongly increases the rate of substrate phosphorylation. Phosphomapping by mass spectrometry indicates that the majority of autophosphorylation sites (37 out of 46) map to a Ser/Thr-rich region immediately N-terminal of the catalytic domain. Deletion of this region prevents substrate phosphorylation without affecting intrinsic catalytic activity suggesting that the Ser/Thr-rich domain contributes to substrate recognition. Surprisingly, the TRPM6-kinase is regulated by an analogous mechanism despite a lack of sequence conservation with the TRPM7 Ser/Thr-rich domain. In conclusion, our findings support a model where massive autophosphorylation outside the catalytic domain of TRPM6 and TRPM7 may facilitate kinase-substrate interactions leading to enhanced phosphorylation of those substrates.Kristopher ClarkJeroen MiddelbeekNick A MorriceCarl G FigdorEdwin LasonderFrank N van LeeuwenPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 3, Iss 3, p e1876 (2008)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kristopher Clark Jeroen Middelbeek Nick A Morrice Carl G Figdor Edwin Lasonder Frank N van Leeuwen Massive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.
description	TRPM6 and TRPM7 are bifunctional proteins expressing a TRP channel fused to an atypical alpha-kinase domain. While the gating properties of TRPM6 and TRPM7 channels have been studied in detail, little is known about the mechanisms regulating kinase activity. Recently, we found that TRPM7 associates with its substrate myosin II via a kinase-dependent mechanism suggesting a role for autophosphorylation in substrate recognition. Here, we demonstrate that the cytosolic C-terminus of TRPM7 undergoes massive autophosphorylation (32+/-4 mol/mol), which strongly increases the rate of substrate phosphorylation. Phosphomapping by mass spectrometry indicates that the majority of autophosphorylation sites (37 out of 46) map to a Ser/Thr-rich region immediately N-terminal of the catalytic domain. Deletion of this region prevents substrate phosphorylation without affecting intrinsic catalytic activity suggesting that the Ser/Thr-rich domain contributes to substrate recognition. Surprisingly, the TRPM6-kinase is regulated by an analogous mechanism despite a lack of sequence conservation with the TRPM7 Ser/Thr-rich domain. In conclusion, our findings support a model where massive autophosphorylation outside the catalytic domain of TRPM6 and TRPM7 may facilitate kinase-substrate interactions leading to enhanced phosphorylation of those substrates.
format	article
author	Kristopher Clark Jeroen Middelbeek Nick A Morrice Carl G Figdor Edwin Lasonder Frank N van Leeuwen
author_facet	Kristopher Clark Jeroen Middelbeek Nick A Morrice Carl G Figdor Edwin Lasonder Frank N van Leeuwen
author_sort	Kristopher Clark
title	Massive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.
title_short	Massive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.
title_full	Massive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.
title_fullStr	Massive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.
title_full_unstemmed	Massive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.
title_sort	massive autophosphorylation of the ser/thr-rich domain controls protein kinase activity of trpm6 and trpm7.
publisher	Public Library of Science (PLoS)
publishDate	2008
url	https://doaj.org/article/7698f9dc761844ae84244a5b619b0d9f
work_keys_str_mv	AT kristopherclark massiveautophosphorylationoftheserthrrichdomaincontrolsproteinkinaseactivityoftrpm6andtrpm7 AT jeroenmiddelbeek massiveautophosphorylationoftheserthrrichdomaincontrolsproteinkinaseactivityoftrpm6andtrpm7 AT nickamorrice massiveautophosphorylationoftheserthrrichdomaincontrolsproteinkinaseactivityoftrpm6andtrpm7 AT carlgfigdor massiveautophosphorylationoftheserthrrichdomaincontrolsproteinkinaseactivityoftrpm6andtrpm7 AT edwinlasonder massiveautophosphorylationoftheserthrrichdomaincontrolsproteinkinaseactivityoftrpm6andtrpm7 AT franknvanleeuwen massiveautophosphorylationoftheserthrrichdomaincontrolsproteinkinaseactivityoftrpm6andtrpm7
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Massive autophosphorylation of the Ser/Thr-rich domain controls protein kinase activity of TRPM6 and TRPM7.

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