Expression and purification of human alpha-7 nicotinic acetylcholine receptor extracellular domain in Pichia pastoris
Issued Date
2023-02-01
Resource Type
ISSN
15131874
Scopus ID
2-s2.0-85152109017
Journal Title
ScienceAsia
Volume
49
Issue
1
Start Page
8
End Page
14
Rights Holder(s)
SCOPUS
Bibliographic Citation
ScienceAsia Vol.49 No.1 (2023) , 8-14
Suggested Citation
Chusuth P., Raksat P., Hannongbua S., Wangkanont K., Ounjai P., Rungrotmongkol T. Expression and purification of human alpha-7 nicotinic acetylcholine receptor extracellular domain in Pichia pastoris. ScienceAsia Vol.49 No.1 (2023) , 8-14. 14. doi:10.2306/scienceasia1513-1874.2022.110 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/82805
Title
Expression and purification of human alpha-7 nicotinic acetylcholine receptor extracellular domain in Pichia pastoris
Author's Affiliation
Other Contributor(s)
Abstract
Human α-7 nicotinic acetylcholine receptor (α-7 nAChR) is an ion channel that plays a significant function in neuronal communication and development. Defects of α-7 nAChR could cause plethora of neurologic and psychiatric impairments such as schizophrenia, Alzheimer’s, and inflammatory disorders. Hence, α-7 nAChR is an interesting candidate for drug development targeting neurological and inflammatory diseases. Therefore, the ability to obtain significant amount of the α-7 nAChR extracellular domain is crucial for further biochemical and structural investigations. We constructed a Pichia pastoris yeast strain that secretes the α-7 nAChR extracellular domain. The recombinant protein was hyperglycosylated and could be deglycosylated with Endoglycosidase H treatment. Cross-linking experiments suggested that the recombinant protein could form oligomeric states, consistent with the known property of the native α-7 nAChR. The optimized expression yield was 4.9 mg/l.