Investigation of SUMO tag on expression, solubility and fibrillation of Alpha synuclein

Saremi, Sabereh; Khajeh, Khosro; Dabirmanesh, Bahareh; Ayyari, Mahdi

Investigation of SUMO tag on expression, solubility and fibrillation of Alpha synuclein

Document Type : Original Research

Authors

Sabereh Saremi

Khosro Khajeh

Bahareh Dabirmanesh

Mahdi Ayyari

Tarbiat Modares University

Abstract

Alpha-synuclein protein (α-syn) is the main factor known in Parkinson's disease. The expression of this protein has challenges. One of these challenges is the presence of protein in bacterial pellet. Studies have shown that the expression of proteins with tags such as small ubiquitin-like modifier (SUMO) increases the expression in the soluble phase, therefore the expression of α-syn with this sequence was investigated to increase the protein in the soluble phase. It has also been shown in studies that SUMOylation has an inhibitory effect on fibrillation, also in this study the effect of the SUMO on alpha-synuclein fibrillation was investigated. The α-syn gene was cloned with SUMO-tag. Nickel sepharose column was used to purify the protein, and dialysis was performed and fibrillation was checked by fluorescence emission of Thioflavin for 72 hours and was observed that the protein with SUMO sequence has a higher expression level, and 95% of the protein is in the soluble phase. On the other hand, it was shown that the SUMO sequence has an inhibitory effect on the process of amyloid fibril formation. The results obtained from previous studies showed that the binding of the SUMO sequence increases the expression and solubility of recombinant proteins. This study revealed that the presence of this sequence contributed to the protein expression level and the protein's presence in the solution phase. On the other hand, observations showed that this sequence has anti-fibrillation properties for proteins with amyloid properties, and in this study showed that SUMO prevents α-syn aggregation.

Keywords

Parkinson’s disease

Alpha synuclein

amyloid fibrils

SUMO-tag

Subjects

Molecular biotechnology

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