Expression and purification of constitutively active S6K1 using dual Bac-to-Bac protein expression system

Abstract

Aim. Since p70S6K1 plays a key role in protein synthesis and cell cycle regulation, it has been implicated in a number of human diseases, such as obesity, diabetes, and cancer. Increased activation of p70S6K1 has been reported in a variety of cancer types, so now it is being investigated as a potential therapeutic target. The purpose of this work was to create a novel more efficient approach of expression and purification of p70S6K1 constitutively active form by using Dual Bac-to-Bac protein expressing system. Conclusions. The current work was focused on the design of a new highly efficient system for the expression and purification of the recombinant His-actS6K1, using pFastBac™ Dual vector co-expressing recombinant GST-PDK1. Recent studies have shown that some cancer types demonstrate the enhanced expression and/or activation of p70S6K, hinting that it may serve as a biomarker for monitoring disease progression. With that in mind, our approach of large-scale expression of His-actS6K1 may offer a rapid, low-cost, and high yield method for producing constitutively active recombinant p70S6K1 and facilitate the development of its potential therapeutic regulators.