title: Comparisons of the conformational stability of cyclin-dependent kinase (CDK) 4-interacting ankyrin repeat (AR) proteins.
abstract: Ankyrin repeat (AR) proteins are one of the most abundant repeat protein classes in nature, and they are involved in numerous physiological processes through mediating protein/protein interactions. The repetitive and modular architecture of these AR proteins may lead to biochemical and biophysical properties distinct from those of globular proteins. It has been demonstrated that like most globular proteins, AR proteins exhibit a two-state, cooperative transition in chemical- and heat-induced unfolding. However, the biophysical characteristics underlying such cooperative unfolding remain to be further investigated. In the present study, we evaluated the conformational stability of a group of cyclin-dependent kinase (CDK) 4-interacting AR proteins, P16, P18, IkappaBalpha, gankyrin, and their truncated mutants under different conditions, including the presence of denaturants, temperature, and pH. Our results showed that the first four N-terminal ARs are required to form a potent and stable CDK4 modulator. Moreover, in spite of their similarities in skeleton structure, CDK4 binding, and cooperative unfolding, P16, P18, IkappaBalpha, and gankyrin exhibited considerably different biophysical properties with regard to the conformational stability, and these differences mainly resulted from the discrepancies in the primary sequence of the relatively conserved AR motifs. Our results also demonstrated that these sequence discrepancies are able to influence the function of AR proteins to a certain extent. Overall, our results provide important insights into understanding the biophysical properties of AR proteins.
authors.affiliation: Ohio State Biochemistry Program, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Guo
authors.forename: Anjali
authors.initials: A
authors.lastname: Mahajan
authors.forename: Chunhua
authors.initials: C
authors.lastname: Yuan
authors.forename: Sang Hoon
authors.initials: SH
authors.lastname: Joo
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Weghorst
authors.forename: Ming-Daw
authors.initials: MD
authors.lastname: Tsai
authors.forename: Junan
authors.initials: J
authors.lastname: Li