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EMBO Rep. 2001 Aug;2(8):703-8. Epub 2001 Jul 19.

FRET-based detection of different conformations of MK2.

Author information

1
Innovationskolleg Zellspezialisierung, Martin-Luther-Universität Halle Wittenberg, 06120 Halle, Germany.

Abstract

MAP kinase-activated protein kinase 2 (MK2 or MAPKAP K2) is a stress-activated enzyme downstream to p38 MAPK. By fusion of green fluorescent protein variants to the N- and C-terminus we analysed conformational changes in the kinase molecule in vitro and in vivo. Activation of MK2 is accompanied by a decrease in fluorescence resonance energy transfer, indicating a transition from an inactive/closed to an active/open conformation with an increase in the apparent distance between the fluorophores of approximately 9 A. The closed conformation exists exclusively in the nucleus. Upon stress, the open conformation of MK2 rapidly becomes detectable in the cytoplasm and accumulates in the nucleus only when Crm1-dependent nuclear export is blocked. Hence, in living cells activation of MK2 and its nuclear export are coupled by a phosphorylation-dependent conformational switch.

PMID:
11463748
PMCID:
PMC1083994
DOI:
10.1093/embo-reports/kve157
[Indexed for MEDLINE]
Free PMC Article

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