中国生物工程杂志

Objective To identify PINK1 interacting with α-synuclein in Parkinson's disease. Methods GST-α-synuclein fusion protein was created by inserting α-synuclein into pGEX-4T-1. The protein was expressed in Escherichia coli and purified on glutathione-Sepharose beads by standard methods. And then incubate with MN9D cells lysates to detect the interaction of PINK1 with α-synuclein. In addition, the full length of PINK1 robustly associated with α-synuclein was also determined by coimmunoprecipitation experiment in MN9D cells. The co-localization of PINK1 with α-synuclein was detected by immunocytochemical staining, and observed by confocal. Results We detected that GST-α-synuclein interact specifically with PINK1, but not GST. Coimmunoprecipitation with an anti-α-synuclein antibody and subsequent Western blotting with an anti-PINK1 antibody confirmed the specific interaction of α-synuclein with PINK1. PINK1 and α-synuclein protein partly co-localize in MN9D cells. Conclusion PINK1 and α-synuclein can interacts with each other and co-localizes partly in MN9D.