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Foldon‐guided self‐assembly of ultra‐stable protein fibers
Author(s) -
Bhardwaj Anshul,
WalkerKopp Nancy,
Wilkens Stephan,
Cingolani Gino
Publication year - 2008
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.036111.108
Subject(s) - denaturation (fissile materials) , biophysics , guanidine , protein engineering , recombinant dna , fusion protein , chimera (genetics) , protein structure , chemistry , protein folding , protein quaternary structure , materials science , crystallography , biochemistry , biology , protein subunit , nuclear chemistry , gene , enzyme
A common objective in protein engineering is the enhancement of the thermodynamic properties of recombinant proteins for possible applications in nanobiotechnology. The performance of proteins can be improved by the rational design of chimeras that contain structural elements with the desired properties, thus resulting in a more effective exploitation of protein folds designed by nature. In this paper, we report the design and characterization of an ultra‐stable self‐refolding protein fiber, which rapidly reassembles in solution after denaturation induced by harsh chemical treatment or high temperature. This engineered protein fiber was constructed on the molecular framework of bacteriophage P22 tail needle gp26, by fusing its helical core to the foldon domain of phage T4 fibritin. Using protein engineering, we rationally permuted the foldon upstream and downstream from the gp26 helical core and characterized gp26‐foldon chimeras by biophysical analysis. Our data demonstrate that one specific protein chimera containing the foldon immediately downstream from the gp26 helical core, gp26(1‐140)‐F, displays the highest thermodynamic and structural stability and refolds spontaneously in solution following denaturation. The gp26‐foldon chimeric fiber remains stable in 6.0 M guanidine hydrochloride, or at 80°C, rapidly refolds after denaturation, and has both N and C termini accessible for chemical/biological modification, thereby representing an ideal platform for the design of self‐assembling nanoblocks.