scholarly journals Beyond Antibodies: Development of a Novel Protein Scaffold Based on Human Chaperonin 10

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Abdulkarim M. Alsultan ◽  
David Y. Chin ◽  
Christopher B. Howard ◽  
Christopher J. de Bakker ◽  
Martina L. Jones ◽  
...  
Keyword(s):  
Protein Scaffold ◽  
Chaperonin 10 ◽  
Novel Protein

Development of a VEGFR-2 antagonist based on a novel protein scaffold (AdNectin)

2005 ◽  
Vol 23 (16_suppl) ◽  
pp. 3150-3150 ◽  
Author(s):  
R. Mamluk ◽  
I. M. Carvajal ◽  
J. M. Bates ◽  
D. M. Kamen ◽  
B. A. Morse ◽  
...  
Keyword(s):  
Protein Scaffold ◽  
Novel Protein

scholarly journals Self-Assembled Amyloid-Like Oligomeric-Cohesin Scaffoldin for Augmented Protein Display on the Saccharomyces cerevisiae Cell Surface

2012 ◽  
Vol 78 (9) ◽  
pp. 3249-3255 ◽  
Author(s):  
Zhenlin Han ◽  
Bei Zhang ◽  
Yi E. Wang ◽  
Yi Y. Zuo ◽  
Wei Wen Su
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Surface ◽  
Fusion Protein ◽  
Fold Increase ◽  
Scaffolding Protein ◽  
Yeast Display ◽  
Protein Scaffold ◽  
Content Type ◽  
Self Assembled ◽  
Novel Protein

ABSTRACTIn this study, a molecular self-assembly strategy to develop a novel protein scaffold for amplifying the extent and variety of proteins displayed on the surface ofSaccharomyces cerevisiaeis presented. The cellulosomal scaffolding protein cohesin and its upstream hydrophilic domain (HD) were genetically fused with the yeast Ure2p N-terminal fibrillogenic domain consisting of residues 1 to 80 (Ure2p1-80). The resulting Ure2p1-80-HD-cohesin fusion protein was successfully expressed inEscherichia colito produce self-assembled supramolecular nanofibrils that serve as a novel protein scaffold displaying multiple copies of functional cohesin domains. The amyloid-like property of the nanofibrils was confirmed via thioflavin T staining and atomic force microscopy. These cohesin nanofibrils attached themselves, via a green fluorescent protein (GFP)-dockerin fusion protein, to the cell surface ofS. cerevisiaeengineered to display a GFP-nanobody. The excess cohesin units on the nanofibrils provide ample sites for binding to dockerin fusion proteins, as exemplified using an mCherry-dockerin fusion protein as well as theClostridium cellulolyticumCelA endoglucanase. More than a 24-fold increase in mCherry fluorescence and an 8-fold increase in CelA activity were noted when the cohesin nanofibril scaffold-mediated yeast display was used, compared to using yeast display with GFP-cohesin that contains only a single copy of cohesin. Self-assembled supramolecular cohesin nanofibrils created by fusion with the yeast Ure2p fibrillogenic domain provide a versatile protein scaffold that expands the utility of yeast cell surface display.

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