title: Isotope labeling of proteins in insect cells.
abstract: Protein targets of contemporary research are often membrane proteins, multiprotein complexes, secreted proteins, or other proteins of human origin. These are difficult to express in the standard expression host used for most nuclear magnetic resonance (NMR) studies, Escherichia coli. Insect cells represent an attractive alternative, since they have become a well-established expression system and simple solutions have been developed for generation of viruses to efficiently introduce the target protein DNA into cells. Insect cells enable production of a larger fraction of the human proteome in a properly folded way than bacteria, as insect cells have a very similar set of cytosolic chaperones and a closely related secretory pathway. Here, the limited and defined glycosylation pattern that insect cells produce is an advantage for structural biology studies. For these reasons, insect cells have been established as the most widely used eukaryotic expression host for crystallographic studies. In the past decade, significant advancements have enabled amino acid type-specific as well as uniform isotope labeling of proteins in insect cells, turning them into an attractive expression host for NMR studies.
authors.affiliation: Novartis Institutes of BioMedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Lukasz
authors.initials: L
authors.lastname: Skora
authors.affiliation: Novartis Institutes of BioMedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Binesh
authors.initials: B
authors.lastname: Shrestha
authors.affiliation: Novartis Institutes of BioMedical Research, Novartis Campus, Basel, Switzerland. Electronic address: alvar.gossert@novartis.com.
authors.forename: Alvar D
authors.initials: AD
authors.lastname: Gossert
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D019476:Insect Proteins / Q000378:metabolism*; D007313:Insecta / Q000166:cytology / Q000378:metabolism*; D007553:Isotope Labeling
chemical_list: D019476:Insect Proteins
keywords: Cell metabolism; Drug discovery; Eukaryotic; Insect cells; Isotope labeling; Membrane proteins; NMR; Protein complex; Selective isotope labeling; Uniform isotope labeling