Journal
BIOORGANIC & MEDICINAL CHEMISTRY
Volume 21, Issue 12, Pages 3504-3510Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.bmc.2013.03.039
Keywords
Protein immobilization; Chemical ligation; Peptide synthesis; Sortase A mediated conjugation; Bioconjugation; Protein modification
Funding
- DFG [TRR67, A4, Be 1264-15/SPP1623]
- Graduate School Leipzig School of Natural Sciences - Building with Molecules and Nanoobjects (BuildMoNa)
- European Union
- Free State of Saxony (ESF)
Ask authors/readers for more resources
Recently, sortase A (SrtA) from Staphyloccus aureus moved into the focus of bioscience because of its ability to incorporate site specific modifications into proteins. The enzyme was mostly used to modify target proteins in an analytical scale, to study biomolecules in their cellular context. In this study, we show the applicability of SrtA mediated ligation for site specific modification of proteins in a large scale. Therefore, the reaction was first optimized using peptides and subsequently new reaction conditions were applied for the large scale biotinylation of interleukin-8. Furthermore, we established C-terminal immobilization of the SrtA on a PEG based resin and could demonstrate maintaining enzymatic activity. Immobilized SrtA significantly facilitates previous ligation protocols as the enzyme can be easily recycled. Also, the removal of excess reaction solution and the whole washing process is significantly accelerated, as centrifugation or filtration techniques can be applied instead of time-consuming chromatography steps. (C) 2013 Elsevier Ltd. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available