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LPS induces pp60 c‐src mediated tyrosine phosphorylation of hsp90 in lung vascular endothelial cells and mouse lung
Author(s) -
Barabutis Nektarios,
Handa Vaishali,
Dimitropoulou Christiana,
Rafikov Ruslan,
Snead Connie,
Kumar Sanjiv,
Joshi Atul,
Thangjam Gagan,
Fulton David,
Black Stephen M,
Patel Vijay,
Catravas John D
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1131.5
Subject(s) - hsp90 , phosphorylation , hsp90 inhibitor , proto oncogene tyrosine protein kinase src , tyrosine phosphorylation , chemistry , tyrosine , microbiology and biotechnology , cancer research , heat shock protein , biology , biochemistry , gene
Post‐translational modifications of Hsp90 may affect client protein binding. We investigated tyrosine (Y) phosphorylation of Hsp90 in inflammation. LPS, a prototypical inflammatory agent, induced concentration‐dependent Y phosphorylation of Hsp90a and Hsp90β in human lung microvascular endothelial cells (HLMVEC). Mass spectroscopy identified a major site of Y phosphorylation as Y309 of Hsp90β (Y300 of Hsp90a). LPS‐induced pY‐Hsp90 was prevented by the Hsp90 inhibitor 17‐AAG in vitro as well as in lungs from LPS‐treated mice, in vivo. Furthermore, 17‐AAG also prevented LPS‐induced pp60 src activation. LPS‐induced pY‐Hsp90 was also prevented by the pp60 src inhibitor, PP2. Additionally, Hsp90 phosphorylation was induced by infecting cells with a constitutively active pp60 src adenovirus, whereas either a dominant‐negative pp60 src adenovirus or reduced expression of pp60 src by a specific siRNA, prevented the LPS‐induced Y phosphorylation of Hsp90. Transfection of HLMVEC with the non‐phosphorylatable Hsp90 Y300F mutant, prevented the LPS‐induced Hsp90 Y phosphorylation and showed a reduced ability to bind the Hsp90 client proteins eNOS and pp60 src . We conclude that inflammatory stimuli cause post‐translational modifications of Hsp90 that are Hsp90‐inhibitor‐sensitive and may be important to the pro‐inflammatory actions of Hsp90. (Supported by HL 06693, HL 093460 and HL 101902).