6Y4J
Engineered Fructosyl Peptide Oxidase
Summary for 6Y4J
| Entry DOI | 10.2210/pdb6y4j/pdb |
| Descriptor | Fructosyl Peptide Oxidase, FLAVIN-ADENINE DINUCLEOTIDE, SULFATE ION, ... (5 entities in total) |
| Functional Keywords | oxidoreductase |
| Biological source | Escherichia coli |
| Total number of polymer chains | 1 |
| Total formula weight | 49337.40 |
| Authors | Donini, S.,Rigoldi, F.,Gautieri, A.,Parisini, E. (deposition date: 2020-02-21, release date: 2021-01-13, Last modification date: 2024-11-13) |
| Primary citation | Rigoldi, F.,Donini, S.,Torretta, A.,Carbone, A.,Redaelli, A.,Bandiera, T.,Parisini, E.,Gautieri, A. Rational backbone redesign of a fructosyl peptide oxidase to widen its active site access tunnel. Biotechnol.Bioeng., 117:3688-3698, 2020 Cited by PubMed Abstract: Fructosyl peptide oxidases (FPOXs) are enzymes currently used in enzymatic assays to measure the concentration of glycated hemoglobin and albumin in blood samples, which serve as biomarkers of diabetes. However, since FPOX are unable to work directly on glycated proteins, current enzymatic assays are based on a preliminary proteolytic digestion of the target proteins. Herein, to improve the speed and costs of the enzymatic assays for diabetes testing, we applied a rational design approach to engineer a novel enzyme with a wider access tunnel to the catalytic site, using a combination of Rosetta design and molecular dynamics simulations. Our final design, L3_35A, shows a significantly wider and shorter access tunnel, resulting from the deletion of five-amino acids lining the gate structures and from a total of 35 point mutations relative to the wild-type (WT) enzyme. Indeed, upon experimental testing, our engineered enzyme shows good structural stability and maintains significant activity relative to the WT. PubMed: 32797625DOI: 10.1002/bit.27535 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.38 Å) |
Structure validation
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