Pleckstrin homology domain of phospholipase C-gamma1 directly binds to 68-kDa neurofilament light chain
Experimental & Molecular Medicine
; : 265-272, 2006.
Article
in En
| WPRIM
| ID: wpr-96564
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WPRO
ABSTRACT
Phosphoinositide-specific phospholipase C-gamma1 (PLC-gamma1) has two pleckstrin homology (PH) domains: an amino-terminal domain (PH1) and a split PH domain (PH2). Here, we show that overlay assay of bovine brain tubulin pool with glutathione-S-transferase (GST)-PLC-gamma1 PH domain fusion proteins, followed by matrix-assisted laser-desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), identified 68-kDa neurofilament light chain (NF-L) as a binding protein of amino-terminal PH domain of PLC-gamma1. NF-L is known as a component of neuronal intermediate filaments, which are responsible for supporting the structure of myelinated axons in neuron. PLC-gamma1 and NF-L colocalized in the neurite in PC12 cells upon nerve growth factor stimulation. In vitro binding assay and immunoprecipitation analysis also showed a specific interaction of both proteins in differentiated PC12 cells. The phosphatidylinositol 4, 5-bisphosphate [PI(4,5)P2] hydrolyzing activity of PLC-gamma1 was slightly decreased in the presence of purified NF-L in vitro, suggesting that NF-L inhibits PLC-gamma1. Our results suggest that PLC-gamma1-associated NF-L sequesters the phospholipid from the PH domain of PLC-gamma1.
Key words
Full text:
1
Index:
WPRIM
Main subject:
Peptides
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Phosphoproteins
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Protein Binding
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Protein Biosynthesis
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Binding Sites
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Molecular Sequence Data
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Blood Proteins
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Amino Acid Sequence
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PC12 Cells
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Neurofilament Proteins
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Experimental & Molecular Medicine
Year:
2006
Type:
Article