Production of selenomethionyl‐derivatized proteins in baculovirus‐infected insect cells
- 1 September 2007
- journal article
- Published by Wiley in Protein Science
- Vol. 16 (9) , 2023-2029
- https://doi.org/10.1110/ps.072931407
Abstract
A protocol is described for the production of both intracellularly expressed and secreted selenomethionyl-derivatized recombinant proteins in baculovirus-infected insect cells. The method results in the production of recombinant soluble proteins with an SeMet occupancy of approximately 75% and with a recovery of approximately 20% that of native protein expression. The method is independent of the percentage methionine content of the protein and is reliable and consistent. Similar results are obtained using either Spodoptera frugiperda Sf9 or Trichoplusia ni High Five insect cells as the expression host, and when cultures are grown in either shake flasks or in Wave BioReactors.Keywords
This publication has 34 references indexed in Scilit:
- Economical parallel protein expression screening and scale-up in Escherichia coliJournal of Structural and Functional Genomics, 2006
- Eukaryotic expression: developments for structural proteomicsActa Crystallographica Section D-Biological Crystallography, 2006
- A time- and cost-efficient system for high-level protein production in mammalian cellsActa Crystallographica Section D-Biological Crystallography, 2006
- Highly efficient selenomethionine labeling of recombinant proteins produced in mammalian cellsProtein Science, 2006
- Crystal structure of human dipeptidyl peptidase IV in complex with a decapeptide reveals details on substrate specificity and tetrahedral intermediate formationProtein Science, 2004
- Structure of a c-Kit Product Complex Reveals the Basis for Kinase TransactivationJournal of Biological Chemistry, 2003
- Crystallization and X-ray analysis of native and selenomethionyl β-mannanase Man5A from blue mussel,Mytilus edulis, expressed inPichia pastorisActa Crystallographica Section D-Biological Crystallography, 2002
- High-Throughput Proteomics: Protein Expression and Purification in the Postgenomic WorldProtein Expression and Purification, 2001
- The expression, characterization, and crystallization of wild-type and selenomethionyl human chorionic gonadotropinEndocrinology, 1995
- Structure of a Fibronectin Type III Domain from Tenascin Phased by MAD Analysis of the Selenomethionyl ProteinScience, 1992