title: High-yield production of functionally active human serum transferrin using a baculovirus expression system, and its structural characterization.
abstract: Recently, there has been much interest in expressing recombinant human serum transferrin (HST) and mutants thereof for structural and functional studies. We have developed a baculovirus expression system for the rapid and efficient production of large quantities of HST (> 20 mg/l). Like native HST, the recombinant protein can bind two ferric ions in the presence of bicarbonate, and is actively taken up by receptor-mediated endocytosis. Secondary structure calculations from CD measurements indicate a content of 42% alpha-helix and 28% beta-sheet. This is the first reported use of a non-mammalian expression system to produce functional HST, and will provide a practical tool to allow expression of a wide range of HST variants for mutagenesis studies.
authors.affiliation: Department of Immunodermatology, Sandoz Research Institute, Vienna, Austria.
authors.forename: S A
authors.initials: SA
authors.lastname: Ali
authors.forename: H C
authors.initials: HC
authors.lastname: Joao
authors.forename: R
authors.initials: R
authors.lastname: Csonga
authors.forename: F
authors.initials: F
authors.lastname: Hammerschmid
authors.forename: A
authors.initials: A
authors.lastname: Steinkasserer
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000737:chemistry; D002460:Cell Line; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007313:Insecta; D010957:Plasmids / Q000378:metabolism; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D014168:Transferrin / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D014168:Transferrin