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Engineering of β-propeller protein scaffolds by multiple gene duplication and fusion of an idealized WD repeat

Paper ID Volume ID Publish Year Pages File Format Full-Text
14107 1148 2006 10 PDF Available
Title
Engineering of β-propeller protein scaffolds by multiple gene duplication and fusion of an idealized WD repeat
Abstract

The ability to design specific amino acid sequences that fold into desired structures is central to engineering novel proteins. Protein design is also a good method to assess our understanding of sequence–structure and structure–function relationships. While β-sheet structures are important elements of protein architecture, it has traditionally been more difficult to design β-proteins than α-helical proteins. Taking advantage of the tandem repeated sequences that form the structural building blocks in a group of β-propeller proteins; we have used a consensus design approach to engineer modular and relatively large scaffolds. An idealized WD repeat was designed from a structure-based sequence alignment with a set of structural guidelines. Using a plasmid sequential ligation strategy, artificial concatemeric genes with up to 10 copies of this idealized repeat were then constructed. Corresponding proteins with 4 through to 10 WD repeats were soluble when over-expressed in Escherichia coli. Notably, they were sufficiently stable in vivo surviving attack from endogenous proteases, and maintained a homogeneous, non-aggregated form in vitro. The results show that the β-propeller scaffold is an attractive platform for future engineering work, particularly in experiments in which directed evolution techniques might improve the stability of the molecules and/or tailor them for a specific function.

Keywords
Protein engineering; Sequence repeats; β-Propeller fold; Consensus design; WD repeats
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Engineering of β-propeller protein scaffolds by multiple gene duplication and fusion of an idealized WD repeat
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biomolecular Engineering - Volume 23, Issue 4, September 2006, Pages 185–194
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