The time-course of agonist-induced solubilization of trimetric Gqα/G11α proteins resolved by two-dimensional electrophoresis

Title:

The time-course of agonist-induced solubilization of trimetric Gqα/G11α proteins resolved by two-dimensional electrophoresis

Creator:

Dana Durchánková, Jiří Novotný, and Petr Svoboda

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:65b024dd-66a6-4457-8517-f917d2191f20
uuid:65b024dd-66a6-4457-8517-f917d2191f20
issn:0862-8408

Subject:

Fyziologie člověka a srovnávací fyziologie, fyziologie, bílkoviny, elektroforéza, thyrotr, physiology, proteins, electrophoresis, G proteins, solubilization, Gqα/G11α, thyrotropin-releasing hormone, TRH receptor, 14, and 612

Type:

article, články, model:article, and TEXT

Format:

print, bez média, and svazek

Description:

Prolonged agonist stimulation results in specific transfer of activated Gα subunits of Gqα/G11α family from particulate membrane fraction to soluble (cytosol) cell fraction isolated as 250 000 x g supernatant. In this study, we have used 2D electrophoresis for more defined resolution of Gα subunits of Gqα/G11α family and followed the time course of solubilization effect. The small signal of soluble G proteins was already detected in control, hormone-unexposed cells. Hormone stimulation resulted in a slow but continuous increase of both intensity and number of immunoreactive signals/spots of these G proteins (10, 30, 60, 120 and 240 min). At longer times of agonist exposure (>2 hours), a marked increase of Gqα/G11α proteins was detected. The maximal level of soluble Gqα/G11α proteins was reached after 16 hours of continuous agonist exposure. At this time interval, eight individual immunoreactive signals of Gqα/G1 α proteins could be resolved. The relative proportion among these spots was 15:42:10:11:7:7:2:5. Solubilization of this class of Gα proteins was thus observed after prolonged agonist stimulation only, induced by ultra high concentration of hormone and in cells expressing a large number of GPCRs. Our data therefore rather indicate tight/persisting binding of Gqα/G11α proteins to the membrane., D. Durchánková, J. Novotný, P. Svoboda., and Obsahuje bibliografii a bibliografické odkazy

Language:

English

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Source:

Physiological research | 2008 Volume:57 | Number:2

Harvested from:

CDK

Metadata only:

false