Pleckstrin homology domain of phospholipase C-gamma1 directly binds to 68-kDa neurofilament light chain.
- Author:
Sung Kuk KIM
1
;
Jang Hyun CHOI
;
Pann Ghill SUH
;
Jong Soo CHANG
Author Information
1. Department of Life Science, College of Natural Science, Daejin University, Kyeonggido 487-711, Korea. jchang@daejin.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
neurofilament protein L;
PC12 cells;
phospholipase C gamma;
phosphatidylinositol 4,5-diphosphate;
protein interaction mapping
- MeSH:
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization;
Rats;
Protein Interaction Mapping;
Protein Biosynthesis/drug effects;
Protein Binding/drug effects;
Phosphoproteins/chemistry/*metabolism;
Phospholipase C gamma/antagonists & inhibitors/chemistry/*metabolism;
Phosphatidylinositol 4,5-Diphosphate/metabolism;
Peptides/chemistry/metabolism;
PC12 Cells;
Neurofilament Proteins/chemistry/*metabolism;
Nerve Growth Factor/pharmacology;
Molecular Weight;
Molecular Sequence Data;
Microtubules/metabolism;
Microscopy, Fluorescence;
Isoenzymes/metabolism/pharmacology/physiology;
Glutathione Transferase/metabolism;
Blotting, Far-Western;
Blood Proteins/chemistry/*metabolism;
Binding Sites;
Animals;
Amino Acid Sequence
- From:Experimental & Molecular Medicine
2006;38(3):265-272
- CountryRepublic of Korea
- Language:English
-
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.
