5h0i

Structure of OaAEP1 asparaginyl peptide ligase in its proenzyme form

spinqualitylabelsall models 5h0i, resolution 2.56Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5h0i is a 2 chain structure with sequence from Oldenlandia affinis. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 2.56Å Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function A0A0N9JZ32_OLDAF Publication Abstract from PubMed Breaking and forming peptidyl bonds are fundamental biochemical reactions in protein chemistry. Unlike proteases that are abundantly available, fast-acting ligases are rare. OaAEP1 is an enzyme isolated from the cyclotide-producing plant oldenlandia affinis that displayed weak peptide cyclase activity, despite having a similar structural fold with other asparaginyl endopeptidases (AEP). Here we report the first atomic structure of OaAEP1, at a resolution of 2.56 A, in its preactivation form. Our structure and biochemical analysis of this enzyme reveals its activation mechanism as well as structural features important for its ligation activity. Importantly, through structure-based mutagenesis of OaAEP1, we obtained an ultrafast variant having hundreds of times faster catalytic kinetics, capable of ligating well-folded protein substrates using only a submicromolar concentration of enzyme. In contrast, the protein-protein ligation activity in the original wild-type OaAEP1 enzyme described previously is extremely weak. Thus, the structure-based engineering of OaAEP1 described here provides a unique and novel recombinant tool that can now be used to conduct various protein labeling and modifications that were extremely challenging before. Engineering a Catalytically Efficient Recombinant Protein Ligase.,Yang R, Wong YH, Nguyen GK, Tam JP, Lescar J, Wu B J Am Chem Soc. 2017 Mar 1. doi: 10.1021/jacs.6b12637. PMID:28199119 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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