Towards the Development of a Recombinant Expression System for Yeast Reticulon Protein

Abstract

Structure determination by X-ray diffraction requires large amounts of purified, functional protein making it especially challenging for membrane proteins due to their low abundance in the cell and instability outside the cell. In this study, we tested the results of our novel screening method for identifying the most promising membrane protein crystallization candidates from native cells through the recombinant expression of the yeast endoplasmic reticulum membrane protein, reticulon(RTN1). Using various mass spectrometry techniques, we were able to determine that RTN1 was not successfully expressed using an E. coli recombinant system, revealing a limitation in the application of our screening method for RTN1.