Alpha-Synuclein Aggregation Assay
Parkinson's disease is a degenerative disease of the nervous system caused by the loss of dopaminergic neurons. The source of this pathological change is still unclear, but studies have shown that Lewy bodies containing alpha-synuclein (αSyn) are the typical neuropathological hallmark of the disease, and almost all patients with Parkinson's disease have different degrees of amyloid deposition in the brain cells, which can directly poison cells and destroy neurotransmitter transmission. Creative Biolabs currently provides professional αSyn in vitro aggregation assays to help your research on neurological diseases.
Fig 1. Schematic structure of a-synuclein. (Stefanis, 2012)
Mechanism of Alpha-Synuclein Aggregation
αSyn is a neuronal protein that is highly enriched in presynaptic nerve terminals. Missense mutations in this protein, gene duplication or triploidy will have a high probability of causing Parkinson's disease. The general consensuses in the field of neuroscience are i) aggregation is the key event in triggering the pathological phenomenon of synuclein; ii) abnormal conformational fibrils formed by aggregation are direct toxicants that affect intracellular targets, disrupt cellular homeostasis and cause neuronal cell death.
In a physiological state, soluble αSyn is naturally an unstructured monomer. After binding to a highly tortuous membrane structure, αSyn undergoes a conformational change and folds into an alpha-helical structure, which is then mediated by its C-terminal region to bind other molecules.
In pathological conditions or high concentrations of αSyn mutants, especially A53T and A30P mutation, the α-helical structure is destroyed, and the β-sheet oligomers named protofibrils are formed. The protofibrils are spherical, circular or string-shaped under electron microscopy and are still initially soluble, but as they combine each other, insoluble fibrils with sizes above 10 nm are emerged. The process was identified as αSyn aggregation, starting from natively unfolded proteins and ending with insoluble fibril formation.
Fig 2. Schematic of the physiological and pathological conformations of αSyn. (Burre, et al., 2018)
Services and Methodology of Alpha-Synuclein Aggregation
Creative Biolabs provides a series of research models including virus-transfected Escherichia coli BL21, LUHMES cell line or Sprague Dawley rats for αSyn aggregation assay. We also induce stable and diverse mutant gene pools in vitro to meet your experimental needs, including A30P, E46K, H50Q, G51D, A53T, and A53E which are commonly used in PD research. It is also feasible to detect αSyn aggregation in vivo or in vitro. In addition to the classical immunological detection methods like western blot, a series of methods based on immunofluorescence or mass spectrometry have also been developed and applied.
Fig 3. Immunohistofluorescence staining in Sprague Dawley rats. (Dimant, et al., 2014)
Although the degree of involvement of αSyn in all Parkinson's cases and the mechanism in neurodegeneration is unclear, the aggregation of αSyn is considered to be an important pathogenic cause of Parkinson's disease. Here, at Creative Biolabs, we can professionally modulate, screen and evaluate αSyn aggregation in vitro. With advanced biotechnology and experienced researchers, we provide comprehensive αSyn aggregation assay services that meet all your experimental design and research needs. Please feel free to contact us for further discussions.
References
- Burré, J.; et al. Cell Biology and Pathophysiology of α-Synuclein. Cold Spring Harbor Perspectives in Medicine. 2018, 8: a024091.
- Dimant, H.; et al. Direct Visualization of CHIP-Mediated Degradation of Alpha-Synuclein InVivo: Implications for PD Therapeutics. PLoS ONE. 2014, 9(3): e92098.
- Stefanis, L. α-Synuclein in Parkinson's disease. Cold Spring Harbor perspectives in medicine. 2012, 2(2): a009399.
