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Lecture 18 -Protein Design and Engineering

Lecture 18 -Protein Design and Engineering - Second Genetic...

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Second Genetic Code : Sequence Structure Folding Design (reverse folding) Computer Algorithm Input Output Folding Sequence Structure Design Structure Sequence
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Protein Stability (thermal) Protein engineering (mutagenesis) 1. S-S bridges a. -CH2-S-S-CH2- b. Analysis of all possibilities (many) c. Energy minimization to reduce to a few plausible candidates d. Site-selective mutations e. Protein synthesis f. Assay: example – T4 lysozyme (x-ray structure known) Reducing degrees of freedom (entropy) increases protein stability
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Protein Stability Cont… 2. Gly and Pro -Gly freedom -Pro Constraints (side chain is fixed by covalent bond to main chain - Gly Pro has propensity to increase stability (more delicate) - Gly Ala usually increase - Pro Ala usually decrease
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Protein Stability Cont… 3. Dipolar stability N-end (-a.a.) C-end (+a.a.) increase stability by mutating residues at N- end of helices from polar to negative (e.g.
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