In vitro assay of neurofilament light chain self-assembly using truncated mutants
- Title
- In vitro assay of neurofilament light chain self-assembly using truncated mutants
- Author
- 이영한
- Keywords
- neurofilament light chain; self-assembly assay; GST pull-down; turbidity; electron microscopy; Western blotting; PC12 cells; SW13 cells
- Issue Date
- 2007-04
- Publisher
- ELSEVIER SCIENCE BV
- Citation
- JOURNAL OF NEUROSCIENCE METHODS, v. 161, No. 2, Page. 199-204
- 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.
- URI
- https://www.sciencedirect.com/science/article/pii/S0165027006005437https://repository.hanyang.ac.kr/handle/20.500.11754/110002
- ISSN
- 0165-0270
- DOI
- 10.1016/j.jneumeth.2006.10.022
- Appears in Collections:
- COLLEGE OF SCIENCE AND CONVERGENCE TECHNOLOGY[E](과학기술융합대학) > ETC
- Files in This Item:
There are no files associated with this item.
- Export
- RIS (EndNote)
- XLS (Excel)
- XML