Ss-stabilizing Proteins Rationally: Intrinsic Disorder-based Design of Stabilizing Disulphide Bridges in GFP
Document Type
Article
Publication Date
2012
Keywords
Green Fluorescent Protein, Differential Scanning Microcalorimetry, Ss-bridge, Ss-bond, Disulphide, Protein Stability, Protein Intrinsic Disorder
Digital Object Identifier (DOI)
https://doi.org/10.1080/07391102.2012.10507414
Abstract
The most attractive and methodologically convenient way to enhance protein stability is via the introduction of disulphide bond(s). However, the effect of the artificially introduced SS-bond on protein stability is often quite unpredictable. This raises the question of how to choose the protein sites in an intelligent manner, so that the ‘fastening’ of these sites by the SS-bond(s) would provide maximal protein stability. We hypothesize that the successful design of a stabilizing SS-bond requires finding highly mobile protein regions. Using GFP as an illustrative example, we demonstrate that the knowledge of the peculiarities of the intramolecular hydrophobic interactions, combined with the understanding of the local intrinsic disorder propensities (that can be evaluated by various disorder predictors, e.g., PONDRFIT), is sufficient to find the candidate sites for the introduction of stabilizing SS-bridge(s). In fact, our analysis revealed that the insertion of the engineered SS-bridge between two highly flexible regions of GFP noticeably increased the conformational stability of this protein toward the thermal and chemical unfolding. Therefore, our study represents a novel approach for the rational design of stabilizing disulphide bridges in proteins.
Was this content written or created while at USF?
Yes
Citation / Publisher Attribution
Journal of Biomolecular Structure and Dynamics, v. 29, issue 4, p. 815-824
Scholar Commons Citation
Melnik, Bogdan S.; Povarnitsyna, Tatiana V.; Glukhov, Anatoly S.; Melnik, Tatiana N.; and Uversky, Vladimir N., "Ss-stabilizing Proteins Rationally: Intrinsic Disorder-based Design of Stabilizing Disulphide Bridges in GFP" (2012). Molecular Medicine Faculty Publications. 494.
https://digitalcommons.usf.edu/mme_facpub/494
