Purification and Functional Characterization of the C-Terminal Domain of the ?-Actin-Binding Protein AIM1 In Vitro.

ABSTRACT: The protein absent in melanoma 1 (AIM1) is a member of the ??-crystal lens superfamily that is associated with the development of multiple cancers. The binding of AIM1 to ?-actin affects the migration and invasion of prostate cancer epithelial cells. The C-terminus of AIM1 is required for the ?-actin interaction. However, the characteristics of AIM1 in vitro and the interaction mode between AIM1 and ?-actin remain unknown. We describe novel methods to prepare pure recombinant AIM1 and identify possible binding modes between AIM1 and ?-actin; we also obtain the crystal of the first two ??-crystallin domains of AIM1 (g1g2) for future structural biology research. We first express and purify AIM1 after cloning the sequence into a modified pET-28a_psp expression vector. Next, we define the minimum unit formed by the ??-crystallin domain repeats that bound to ?-actin and perform its physiological function. Finally, we made the structural model of the AIM1 g1g2 that can be used to guide future biomedical investigations and prostate cancer research.

SUBMITTER: Wu F 

PROVIDER: S-EPMC6320856 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature