Abstract
Myocyte enhancer factor 2 (MEF2) proteins play a pivotal role in the differentiation of cardiac and skeletal muscle cells. MEF2 factors are regulated by histone deacetylase enzymes such as histone deacetylase 5 (HDAC5). HDAC5 in turn is responsive to Ca(2+) signaling mediated by the intracellular calcium sensor calmodulin. Here a combination of proteolytic fragmentation, matrix-assisted laser desorption ionization mass spectrometry, Edman degradation, circular dichroism, gel filtration, and surface plasmon resonance studies is utilized to define and characterize a stable core domain of HDAC5 and to examine its interactions with MEF2a and calmodulin. Results from real time binding experiments provide evidence for direct interaction of Ca(2+)/calmodulin with HDAC5 inhibiting MEF2a association with this enzyme.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Motifs
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Amino Acid Sequence
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Calcium / metabolism*
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Calmodulin / chemistry
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Calmodulin / metabolism*
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Cells, Cultured
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Chromatography, Gel
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Circular Dichroism
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DNA-Binding Proteins / antagonists & inhibitors
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DNA-Binding Proteins / metabolism*
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Egtazic Acid / pharmacology
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Histone Deacetylase Inhibitors*
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Histone Deacetylases / metabolism*
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Humans
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Kinetics
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MADS Domain Proteins
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MEF2 Transcription Factors
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Mass Spectrometry
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Models, Molecular
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Molecular Sequence Data
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Mutation
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Myogenic Regulatory Factors
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Promoter Regions, Genetic
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Protein Binding
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Protein Structure, Tertiary
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Sequence Homology, Amino Acid
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Surface Plasmon Resonance
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Time Factors
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Transcription Factors / antagonists & inhibitors
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Transcription Factors / metabolism*
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Trypsin / pharmacology
Substances
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Calmodulin
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DNA-Binding Proteins
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Histone Deacetylase Inhibitors
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MADS Domain Proteins
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MEF2 Transcription Factors
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MEF2A protein, human
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Myogenic Regulatory Factors
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Transcription Factors
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Egtazic Acid
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Trypsin
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HDAC5 protein, human
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Histone Deacetylases
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Calcium