8z2j

Substrate analog a012 bound form of PET-degrading cutinase mutant Cut190**SS_S176A

spinqualitylabelsall models 8z2j, resolution 1.89Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 8z2j is a 2 chain structure with sequence from Saccharomonospora viridis. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.89Å Ligands: A1L0M, ACT, CA, CL, MPD Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function W0TJ64_9PSEU Publication Abstract from PubMed We previously revealed the structural basis of Ca(2+) dependent regulation of a polyethylene terephthalate (PET)-degrading enzyme, Cut190, and proposed a unique reaction cycle in which the enzyme repeatedly binds and releases Ca(2+). Here, we report crystal structures of Cut190 mutants with high thermal stability complexed with PET-like ligands that contain aromatic rings. The structural information has allowed us to perform further computational analyses using a PET-trimer bound model. Our multicanonical molecular dynamics simulations and subsequent analyses of the free energy landscapes revealed a novel intermediate form that occurs during the enzymatic reaction cycle. Furthermore, the computational analyses were used to investigate the effect of the point mutations F77L and F81L in the Ca(2+)-binding site, which showed that the former stabilizes the engaged and open forms to improve transition between the open and active forms, while the latter extremely increases the open form. Subsequent experiments showed that the F77L mutation increased the activity, while the F81L mutation decreased the activity. Our computational analysis has enabled us to explore the dynamics of Cut190 on a completely new level, providing key insights into how the balance between the various conformations influences the reaction cycle and ultimately how to improve the reaction cycle. Structural dynamics of the Ca(2+)-regulated cutinase towards structure-based improvement of PET degradation activity.,Numoto N, Kondo F, Bekker GJ, Liao Z, Yamashita M, Iida A, Ito N, Kamiya N, Oda M Int J Biol Macromol. 2024 Oct 15;281(Pt 4):136597. doi: , 10.1016/j.ijbiomac.2024.136597. PMID:39419144 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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