A Protein Expression Toolkit for Studying Signaling in T Cells

Methods Mol Biol. 2017:1584:451-472. doi: 10.1007/978-1-4939-6881-7_28.

Abstract

Innate and adaptive cellular immunity is dependent on interactions of cell surface receptors that initiate signaling, resulting in the formation of the immunological synapse and targeted delivery of effector functions. There has been considerable interest over the past 30 years in methods for isolating the extracellular regions of these receptors and components of the cytoplasmic signaling networks. This chapter describes our current protein expression toolkit used for structural studies of signaling proteins and the functional reconstitution of model cell surfaces, which comprises both bacterial and mammalian cell-based protein expression methodologies.

Keywords: Adhesion; Affinity; Chromatography; Glycosylation; Inclusion bodies; Protein folding; Receptors.

MeSH terms

  • Animals
  • Gene Expression*
  • Humans
  • Recombinant Proteins* / biosynthesis
  • Recombinant Proteins* / genetics
  • Recombinant Proteins* / immunology
  • Signal Transduction* / genetics
  • Signal Transduction* / immunology

Substances

  • Recombinant Proteins