Heterochiral Knottin Protein: Folding and Solution Structure
Year: 2017
Type: article
Abstract: Homochirality is a general feature of biological macromolecules, and Nature includes few examples of heterochiral proteins. Herein, we report on the design, chemical synthesis, and structural characterization of heterochiral proteins possessing loops of amino acids of chirality opposite to that of the rest of a protein scaffold. Using the protein Ecballium elaterium trypsin inhibitor II, we discover that selective β-alanine substitution favors the efficient folding of our heterochiral constructs. Solution nuclear magnetic resonance spectroscopy of one such heterochiral protein reveals a homogeneous global fold. Additionally, steered molecular dynamics simulation indicate β-alanine reduces the free energy required to fold the protein. We also find these heterochiral proteins to be more resistant to proteolysis than homochiral l-proteins. This work informs the design of heterochiral protein architectures containing stretches of both d- and l-amino acids.
Source: Biochemistry
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: WIDE-BORE 9.4 TESLA NMR SPECTROMETER FOR UW-MADISON, Upgrade of the NMRFAM 750 MHz NMR Spectrometer, Interdepartmental Biotechnology Training Program, National Biomedical NMR Resource at Madison, Integrated state-of-the-art LC-MS/NMR system for the University of Wisconsin, National Biomedical NMR Resource at Madison, CONSOLE UPGRADES FOR NMR FACILITY, NMR Spectrometer System Optimized for Direct 13C or 15N Detection, G7988586476, Cysteine Arylation
OpenAlex ID: W2759093161