title: Production of recombinant serpins in Escherichia coli.
abstract: Serpins represent a diverse family of proteins that are found in a wide range of organisms and cellular locations. In order to study them, most need to be produced recombinantly, as isolation from their source is not always possible. Due to their relatively uncomplicated structure (single domain, few posttranslational modifications), the serpins are usually amenable to expression in Escherichia coli, which offers a fast and cost-effective solution for the generation of large amounts of protein. This chapter outlines the general procedures used in the expression and subsequent purification of serpins in E. coli, with a particular focus on the methods used for antitrypsin, the archetypal member of the family.
authors.affiliation: Department of Biochemistry, Monash University, Clayton, Australia.
authors.forename: Mary C
authors.initials: MC
authors.lastname: Pearce
authors.forename: Lisa D
authors.initials: LD
authors.lastname: Cabrita
mesh_terms: D002250:Carbon Radioisotopes / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D003001:Cloning, Molecular / Q000379:methods*; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000145:classification / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011401:Promoter Regions, Genetic; D017510:Protein Folding; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D012995:Solubility; D013194:Staining and Labeling / Q000379:methods*; D014169:Transformation, Bacterial; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D002250:Carbon Radioisotopes; D003470:Culture Media; D011994:Recombinant Proteins; D000515:alpha 1-Antitrypsin