Conformational Stabilization and Rapid Labeling of a 29-Residue Peptide by a Small Molecule Reaction Partner
| Autor: | Zachary P. Gates, Alexander J. Mijalis, Zhen-Yu J. Sun, Bradley L. Pentelute, Ethan D. Evans |
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| Rok vydání: | 2019 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences Chemistry Stereochemistry Protein Conformation 030302 biochemistry & molecular biology Peptide Biochemistry Small molecule Orders of magnitude (mass) Reaction rate 03 medical and health sciences Residue (chemistry) Reaction rate constant Enzyme Stability Mutation Reactivity (chemistry) Computer Simulation Amino Acid Sequence Cysteine Amino Acids Peptides Protein Binding |
| Zdroj: | Biochemistry. 58(10) |
| ISSN: | 1520-4995 |
| Popis: | A 29-residue peptide (MP01), identified by in vitro selection for reactivity with a small molecule perfluoroaromatic, was modified and characterized using experimental and computational techniques, with the goal of understanding the molecular basis of its reactivity. These studies identified a six-amino acid point mutant (MP01-Gen4) that exhibited a reaction rate constant of 25.8 ± 1.8 M–1 s–1 at pH 7.4 and room temperature, approximately 2 orders of magnitude greater than that of its progenitor sequence and 3 orders of magnitude greater than background cysteine reactivity. MP01-Gen4 appeared to be conformationally dynamic and exhibited several properties reminiscent of larger protein molecules, including denaturant-sensitive structure and reactivity. We believe the majority of the reaction rate enhancement can be attributed to interaction of MP01-Gen4 with the perfluoroaromatic probe, which was found to stabilize a helical conformation of both MP01-Gen4 and nonreactive Cys-to-Ser or Cys-to-Ala variants. ... |
| Databáze: | OpenAIRE |
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