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Site-specific tetrameric streptavidin-protein conjugation using sortase A

Paper ID Volume ID Publish Year Pages File Format Full-Text
24102 43497 2011 6 PDF Available
Title
Site-specific tetrameric streptavidin-protein conjugation using sortase A
Abstract

Streptavidin is tetrameric protein which has tight and specific biotin binding affinity, and streptavidin modification of proteins or small molecules is widely used for biotechnology tool. Here, we demonstrate site-specific streptavidin-protein conjugation using enzymes. We focused on sortase A, a transpeptidase from Staphylococcus aureus. A streptavidin-tagged LPETG motif (Stav-LPETG) was expressed in Escherichia coli. We achieved soluble streptavidin expression in E. coli without refolding using a cold shock expression system. Then we successfully conjugated Stav-LPETG with pentaglycine-appended green fluorescence protein (Gly5-GFP) or triglycine-appended glucose oxidase (Gly3-GOD) using sortase A. SDS-PAGE analysis showed site-specific tetrameric streptavidin-protein conjugation with the tagged proteins. In addition, the functions of a Stav-GOD conjugate, i.e., biotin-binding and glucose oxidase activity, were significantly higher compared to those of streptavidin-GOD conjugates prepared by chemical modification.

Keywords
Streptavidin; Sortase; Modification; Cold shock expression; Site-specific
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Site-specific tetrameric streptavidin-protein conjugation using sortase A
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 152, Issues 1–2, 10 March 2011, Pages 37–42
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us