Leishmania tarentolae for the Production of Multi-subunit Complexes.

Abstract

Multi-subunit protein complexes are involved in a wide variety of cellular processes including DNA replication, transcriptional regulation, signal transduction, protein folding and degradation. A better understanding of the function of these protein complexes requires structural insights into the molecular arrangement and interactions of their constituent subunits. However, biochemical and structural analysis of multi-subunit protein complexes is still limited because of technical difficulties with their recombinant expression and reconstitution. This chapter presents an overview of a novel protein expression system based on Leishmania tarentolae, a unicellular protozoan parasite of lizards, and practical considerations for the production of multi-subunit protein complexes. The Leishmania tarentolae expression system offers fully eukaryotic protein expression with post-translational modifications but with ease of handling similar to bacteria. This chapter also summarizes studies on the production of laminins, large heterotrimeric glycoproteins of the extracellular matrix, using this expression system. In addition, a recently developed Leishmania tarentolae-based cell-free translation system is briefly described.