title: A Protein Expression Toolkit for Studying Signaling in T Cells.
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.
authors.affiliation: Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine and MRC Human Immunology Unit, John Radcliffe Hospital, University of Oxford, Headington, Oxford, OX3 9DS, UK.
authors.forename: Ana Mafalda
authors.initials: AM
authors.lastname: Santos
authors.affiliation: Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine and MRC Human Immunology Unit, John Radcliffe Hospital, University of Oxford, Headington, Oxford, OX3 9DS, UK.
authors.forename: Jiandong
authors.initials: J
authors.lastname: Huo
authors.affiliation: Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine and MRC Human Immunology Unit, John Radcliffe Hospital, University of Oxford, Headington, Oxford, OX3 9DS, UK.
authors.forename: Deborah
authors.initials: D
authors.lastname: Hatherley
authors.affiliation: Department of Structural Biology, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, 862-0973, Japan.
authors.forename: Mami
authors.initials: M
authors.lastname: Chirifu
authors.affiliation: Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine and MRC Human Immunology Unit, John Radcliffe Hospital, University of Oxford, Headington, Oxford, OX3 9DS, UK. simon.davis@imm.ox.ac.uk.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Davis
mesh_terms: D000818:Animals; D015870:Gene Expression; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology
chemical_list: D011994:Recombinant Proteins
keywords: Adhesion; Affinity; Chromatography; Glycosylation; Inclusion bodies; Protein folding; Receptors