Please use this identifier to cite or link to this item:
|Title:||Versatile Protein Biotinylation Strategies for Potential High-Throughput Proteomics|
|Source:||Lue, R.Y.P., Chen, G.Y.J., Hu, Y., Zhu, Q., Yao, S.Q. (2004-02-04). Versatile Protein Biotinylation Strategies for Potential High-Throughput Proteomics. Journal of the American Chemical Society 126 (4) : 1055-1062. ScholarBank@NUS Repository. https://doi.org/10.1021/ja037914g|
|Abstract:||We present intein-mediated approaches for efficient biotinylation of proteins site-specifically. The reactive C-terminal thioester generated from intein-assisted protein splicing (either in vitro or in live cells) served as an attractive and exclusive site for attaching cysteine-containing biotin. Using these novel biotinylation strategies, we were able to efficiently biotinylate many proteins from different biological sources in a potentially high-throughput, high-content fashion. Some of these proteins were subsequently immobilized, in a very simple manner, onto different avidin-functionalized solid surfaces for applications such as protein microarray and surface plasmon resonance (SPR) spectroscopy, highlighting the numerous advantages of using biotin over other tags (e.g., GST, His-tag, etc.) as the method of choice in protein purification/immobilization. In addition, our intein-mediated strategies provided critical advantages over other protein biotinylation strategies in a number of ways. For the first time, we also successfully demonstrated that intein-mediated protein biotinylation proceeded adequately inside both bacterial and mammalian living cells, as well as in a cell-free protein synthesis system. Taken together, our results indicate the versatility of these intein-mediated strategies for potential high-throughput proteomics applications. They may also serve as useful tools for various biochemical and biophysical studies of proteins both in vitro and in vivo.|
|Source Title:||Journal of the American Chemical Society|
|Appears in Collections:||Staff Publications|
Show full item record
Files in This Item:
There are no files associated with this item.
checked on Mar 21, 2018
WEB OF SCIENCETM
checked on Mar 21, 2018
checked on Mar 11, 2018
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.