Investigation of purified Survivin interaction with Caspase ۹ using Native-PAGE
سال انتشار: 1402
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 31
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شناسه ملی سند علمی:
BCSCD02_007
تاریخ نمایه سازی: 11 فروردین 1403
چکیده مقاله:
Background: Survivin is one of the smallest members of the IAP family. Improper expression of this protein is found in much human cancer, which inhibits caspases and creates resistance to therapies. Survivin protein is shown as a cancer marker due to its expression is higher in cancer cells compared to normal tissues. Since IAP proteins have the potential to interact with caspase ۹ and so for no interaction between survivin and caspase ۹ has been investigated by purified proteins. In this study, the interaction of these proteins was analyzed using Native-PAGE gel. Methods: The vectors carrying the caspase ۹ wild-type and Survivn genes from non-expressive DH۵ α bacteria were extracted separately, and transferred independently to the BL۲۱(DE۳) expression strain. Both proteins were expressed by IPTG and lactose inducers in ۲xYT medium and then purified. Two proteins were mixed t ۱:۱ ۲۲ ۴ ۵۲ C and cross-linked by glutaraldehyde. This process was stopped by Tris ۱M and estimated by Native-PAGE gel.Results: The results of SDS-PAGE gel showed that a significant amounts of both purified proteins were obtained. It was also observed from the result of Native-PAGE gel that survivin protein interacted with caspase ۹ as XIAP-BIR۳ . Conclusion: Because survivin has an alpha helix instead of a RING-finger domain, it binds zinc ions to the BIR domain. Native-PAGE gel showed survivin interacted with caspase ۹ . Adding a chelating agent like EDTA can be found whether the detach zinc ion from survivin destroy the possible interaction with caspase ۹ or not, which needs to be investigated.
کلیدواژه ها:
نویسندگان
Mahsa Tirmomenin
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
Farangis Ataei
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
Saman Hosseinkhani
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran