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A temperature-dependent conformational shift in p38α MAP kinase substrate binding region associated with changes in substrate phosphorylation profile.

J. Biol. Chem.. 2019; 
DeredgeDaniel,WintrodePatrick L,TulapurkarMohan E,NagarsekarAshish,ZhangYingua,WeberDavid J,ShapiroPaul,HasdayJeffr
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Bacterial Expression System … to create pETDuet-p38β. The coding sequences for ATF2 and MK2 were optimized for bacterial expression, synthesized (Genscript; Piscataway, NH) and ligated between the BamHI and HindIII sites in pRSetA. The p38α and … Get A Quote

摘要

Febrile-range hyperthermia (FRH) worsens and hypothermia mitigates lung injury and temperature dependence of lung injury is blunted by inhibitors of p38 mitogen-activated protein kinase (MAPK). Of the two predominant p38 isoforms, p38α is proinflammatory and p38β is cytoprotective. Here, we analyzed the temperature dependence of p38 MAPK activation, substrate interaction, and tertiary structure. Incubating HeLa cells at 39.5°C stimulated modest p38 activation, but did not alter tumor necrosis factor-α (TNFα)-induced p38 activation. In in vitro kinase (IVK) assays containing activated p38α and MAPK-activated kinase-2 (MK2), MK2 phosphorylation was 14.5-fold greater at 39.5°C than 33°C. By com... More

关键词

MAPKAPK-2,hydrogen-deuterium exchange,hyperthermia,p38 MAPK,protein structure,pyrogen,signaling,substrate specificity,substrate-docking site,temperature-depend