title: Enzyme-specific activation versus leaving group ability.
abstract: Enzyme-specific activation and the substrate mimetics strategy are effective ways to circumvent the limited substrate recognition often encountered in protease-catalyzed peptide synthesis. A key structural element in both approaches is the guanidinophenyl (OGp) ester, which enables important interactions for affinity and recognition by the enzyme--at least, this is usually the explanation given for its successful application. In this study we show that leaving group ability is of equal or even greater importance. To this end we used both experimental and computational methods: 1) synthesis of close analogues of OGp, and their evaluation in a dipeptide synthesis assay with trypsin, 2) molecular docking studies to provide insights into the binding mode, and 3) ab initio calculations to evaluate their electronic properties.
authors: Institute for Molecules and Materials, Radboud University Nijmegen, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
authors: Roseri J A C
authors: RJ
authors: de Beer
authors: Berry
authors: B
authors: Bögels
authors: Gijs
authors: G
authors: Schaftenaar
authors: Barbara
authors: B
authors: Zarzycka
authors: Peter J L M
authors: PJ
authors: Quaedflieg
authors: Floris L
authors: FL
authors: van Delft
authors: Sander B
authors: SB
authors: Nabuurs
authors: Floris P J T
authors: FP
authors: Rutjes
mesh_terms: D055162:Biocatalysis; D001681:Biological Assay; D004151:Dipeptides / Q000138:chemical synthesis*; D004789:Enzyme Activation; D004952:Esters; D006860:Hydrogen Bonding; D006868:Hydrolysis; D008958:Models, Molecular; D018716:Molecular Mimicry; D011487:Protein Conformation; D011789:Quantum Theory; D012996:Solutions; D013379:Substrate Specificity; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004151:Dipeptides; D004952:Esters; D012996:Solutions; D014357:Trypsin