Mutations of G38R and D40G cause amyotrophic lateral sclerosis by reducing Annexin A11 protein stability.
10.11817/j.issn.1672-7347.2018.06.001
- Author:
Di LIAO
1
;
Qiao LIAO
1
;
Cao HUANG
2
;
Fangfang BI
1
Author Information
1. Department of Neurology, Xiangya Hospital, Central South University, Changsha 410008, China.
2. Department of Pathology, Anatomy & Cell Biochemistry, Thomas Jefferson University, Philadelphia PA 19107, USA.
- Publication Type:Journal Article
- MeSH:
Amyotrophic Lateral Sclerosis;
genetics;
metabolism;
Annexins;
chemistry;
genetics;
metabolism;
HEK293 Cells;
Humans;
Mutation;
Plasmids;
genetics;
Protein Stability;
Solubility;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2018;43(6):577-582
- CountryChina
- Language:Chinese
-
Abstract:
To explore the role of the mutations G38R and D40G of Annexin A11 (ANXA11) in the onset of amyotrophic lateral sclerosis (ALS).
Methods: The plasmids expressing ANXA11 wild type protein, ANXA11 G38R protein and ANXA11 D40G protein were constructed, respectively. The recombinant plasmids were then transfected into HEK293 cells respectively followed by cycloheximide (CHX) treatment for 0, 2, 4 and 8 h. The protein expressions of ANXA11 wild type, ANXA11 G38R and ANXA11 D40G mutations were determined by Western blot. Gray analysis by Image J was performed to compare the half-life of each protein. The NSC-34 cell lines constantly expressing ANXA11 wild type protein, ANXA11 G38R protein and ANXA11 D40G protein were established. The cells were treated with NP-40 lysis buffer to examine the protein solubility by Western blot.
Results: Both ANXA11 G38R protein and ANXA11 D40G protein showed a shorter half-life than ANXA11 wild type protein (P<0.05), while there was no difference between ANXA11 G38R protein and ANXA11 D40G protein (P>0.05). There was no visible insoluble substance in the NP-40 lysates for ANXA11 wild type protein, ANXA11 G38R protein and ANXA11 D40G protein.
Conclusion: G38R and D40G mutations reduce the stability of ANXA11 protein. G38R and D40G mutations do not alter ANXA11 solubility.