In vitro assay of neurofilament light chain self-assembly using truncated mutants
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SCOPUS
- Title
- In vitro assay of neurofilament light chain self-assembly using truncated mutants
- Authors
- Kim, SK; Cho, SM; Lee, IB; Lee, YH; Kang, JH; Choi, JH; Suh, PG; Chang, JS
- Date Issued
- 2007-04-15
- Publisher
- ELSEVIER SCIENCE BV
- Abstract
- Neurofilaments (NFs) are heteropolymers composed of light (NF-L), middle (NF-M), and heavy (NF-H) subunits, present in most neurons. NF-L polymerizes on its own to provide a scaffold on which regular NFs form via the cross-bridging of NF-M or NF-H. To clarify the mechanism of regulation of NF-L self-assembly, we developed an assay using truncated mutant NF-L fused to glutathione-S transferase (GST). Western immunoblotting data show that the GST-fused head-rod domains of NF-L are necessary and sufficient for detecting assembled NF-L. The levels of self-assembled NF-L subunits detected using GST fusion proteins were consistent with those detected by electron microscopy and turbidity assay. Our results collectively imply that GST-fused head-rod domains of NF-L are critical tools for analyzing NF-L self-assembly in vitro. (C) 2006 Elsevier B.V. All rights reserved.
- Keywords
- neurofilament light chain; self-assembly assay; GST pull-down; turbidity; electron microscopy; Western blotting; PC12 cells; SW13 cells; AMYOTROPHIC-LATERAL-SCLEROSIS; PROTEIN-KINASE; INTERMEDIATE-FILAMENTS; AXONAL-TRANSPORT; END DOMAINS; ROD DOMAIN; NF-M; DISEASE; PHOSPHORYLATION; CELLS
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/23420
- DOI
- 10.1016/J.JNEUMETH.2
- ISSN
- 0165-0270
- Article Type
- Article
- Citation
- JOURNAL OF NEUROSCIENCE METHODS, vol. 161, no. 2, page. 199 - 204, 2007-04-15
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