Expression and purification of active human kinases using Pichia pastoris as a general-purpose host.

ABSTRACT:

Background

The heterologous expression of human kinases in good purity and in a monomeric, soluble and active form can be challenging. Most of the reported successful attempts are carried out in insect cells as a host. The use of E. coli for expression is limited to a few kinases and usually is facilitated by large solubility tags that can limit biophysical studies and affect protein-protein interactions. In this report, we evaluate the methylotrophic yeast Pichia pastoris (P. pastoris) as a general-purpose host for expression of human kinases.

Methods

Six diverse kinases were chosen due to their therapeutic importance in human cancers. Tested proteins include serine/threonine kinases cyclin-dependent kinases 4 and 6 (CDK4 and 6) and aurora kinase A (AurKA), receptor tyrosin

SUBMITTER: Abdel Aziz MH 

PROVIDER: S-EPMC11655031 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature