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Isolation and Characterization of Histone Deacetylase Kinases from Failing Human Heart
Author(s) -
Huynh Khai Quang,
Hood David B,
Mckinsey Timothy A
Publication year - 2009
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.23.1_supplement.486.2
Subject(s) - histone deacetylase 5 , camk , kinase , biology , protein kinase a , microbiology and biotechnology , histone deacetylase , hdac4 , histone , biochemistry , autophosphorylation , gene
Histone deacetylase 5 (HDAC5) represses pathological cardiac gene expression through interactions with pro‐hypertrophic transcription factors. In response to stress signals, HDAC5 is phosphorylated and exported to the cytoplasm, resulting in derepression of downstream target genes that promote pathological cardiac remodeling. Overexpression and in vitro studies have identified multiple putative HDAC5 kinases, most of which are members of the Ca 2+ /calmodulin‐dependent protein kinase (CaMK) superfamily (e.g., protein kinase D (PKD), microtubule associated kinase (MARK) and CaMK). We employed an unbiased approach to identify endogenous HDAC5 kinases from human heart. A six‐step biochemical purification followed by mass spectrometry uncovered three HDAC5 kinases in the myocardium of patients with end‐stage heart failure: PKD1, PKD2 and MARK3. Kinetic studies with recombinant enzymes and HDAC5 substrate in vitro confirmed the capacity of multiple members of the CaMK superfamily to phosphorylate HDAC5, including checkpoint kinase (CHK) 1/2, salt inducible kinase (SIK) 1, mitogen‐ and stress‐activated kinase (MSK) 1/2, death‐associated protein kinase (DAPK) 1, testis‐specific serine/threonine kinase (TSSK) 1 and PIM‐1. However, only PKDs exhibited low K m values and high catalytic activity (k cat /K m ) toward HDAC5, and co‐immunoprecipitation studies revealed selective physical association of HDAC5 with PKD. These findings support a role for PKD isoforms in the control of pathological cardiac remodeling through their neutralizing effects on HDAC5.