The role of streptavidin and its variants in catalysis by biotinylated secondary amines
| Autor: | Louis C. Morrill, Katarzyna Świderek, Nicolò Santi, Raquel Castillo, Magdalena Lipka-Lloyd, Alexander R. Nödling, Louis Yp Luk, Vicent Moliner, Yi Jin |
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| Rok vydání: | 2021 |
| Předmět: |
Streptavidin
Addition reaction catalysis Stereochemistry Chemistry Organic Chemistry Iminium Protein engineering ResearchInstitutes_Networks_Beacons/manchester_institute_of_biotechnology Biochemistry Residue (chemistry) chemistry.chemical_compound Protein structure Biotinylation Manchester Institute of Biotechnology biotinylated secondary amines Protein quaternary structure Physical and Theoretical Chemistry |
| Zdroj: | Repositori Universitat Jaume I Universitat Jaume I Organic & Biomolecular Chemistry Nödling, A R, Santi, N, Castillo, R, Lipka-Lloyd, M, Jin, Y, Morrill, L C, Świderek, K, Moliner, V & Luk, L Y P 2021, ' The role of streptavidin and its variants in catalysis by biotinylated secondary amines ', Organic & biomolecular chemistry, vol. 19, no. 47, pp. 10424-10431 . https://doi.org/10.1039/d1ob01947c |
| DOI: | 10.1039/D1OB01947C |
| Popis: | Here, we combine the use of host screening, protein crystallography and QM/MM molecular dynamics simulations to investigate how the protein structure affects iminium catalysis by biotinylated secondary amines in a model 1,4 conjugate addition reaction. Monomeric streptavidin (M-Sav) lacks a quaternary structure and the solvent-exposed reaction site resulted in poor product conversion in the model reaction with low enantio- and regioselectivities. These parameters were much improved when the tetrameric host T-Sav was used; indeed, residues at the symmetrical subunit interface were proven to be critical for catalysis through a mutagenesis study. The use of QM/MM simulations and the asymmetric dimeric variant D-Sav revealed that both Lys121 residues which are located in the hosting and neighboring subunits play a critical role in controlling the stereoselectivity and reactivity. Lastly, the D-Sav template, though providing a lower conversion than that of the symmetric tetrameric counterpart, is likely a better starting point for future protein engineering because each surrounding residue within the asymmetric scaffold can be refined for secondary amine catalysis. Here, we combine the use of host screening, protein crystallography and QM/MM molecular dynamics simulations to investigate how protein enviroment affects iminium catalysis by biotinylated secondary amines in a model 1,4 conjugate addition reaction. |
| Databáze: | OpenAIRE |
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