Said M Afify, Ghmkin Hassan, Hend M Nawara, Maram H Zahra, Yanning Xu, Md Jahangir Alam, Koichi Saitoh, Hager Mansour, Hagar A Abu Quora, Mona Sheta, Sadia Monzur, Juan Du, Sue Young Oh, Akimasa Seno, David S Salomon, Masaharu Seno
Human Cripto-1 is a member of the epidermal growth factor (EGF)-Cripto-FRL-1-Cryptic (CFC) family family and performs critical roles in cancer and various pathological and developmental processes. Recently we demonstrated that a soluble form of Cripto-1 suppresses the self-renewal and enhances the differentiation of cancer stem cells (CSCs). A functional form of soluble Cripto-1 was found to be difficult to obtain because of the 12 cysteine residues in the protein which impairs the folding process. Here, we optimized the protocol for a T7 expression system, purification from inclusion bodies under denatured conditions refolding of a His-tagged Cripto-1 protein...
July 2022: Journal of Cellular Biochemistry