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Full-Text Articles in Physical Sciences and Mathematics
Molecular Recognition Of Insulin By A Synthetic Receptor, Jordan M. Chinai, Alexander B. Taylor, Lisa M. Ryno, Nicholas D. Hargreaves, Christopher A. Morris, P John Hart, Adam R. Urbach
Molecular Recognition Of Insulin By A Synthetic Receptor, Jordan M. Chinai, Alexander B. Taylor, Lisa M. Ryno, Nicholas D. Hargreaves, Christopher A. Morris, P John Hart, Adam R. Urbach
Chemistry Faculty Research
The discovery of molecules that bind tightly and selectively to desired proteins continues to drive innovation at the interface of chemistry and biology. This paper describes the binding of human insulin by the synthetic receptor cucurbit[7]uril (Q7) in vitro. Isothermal titration calorimetry and fluorescence spectroscopy experiments show that Q7 binds to insulin with an equilibrium association constant of 1.5 × 106 M−1 and with 50−100-fold selectivity versus proteins that are much larger but lack an N-terminal aromatic residue, and with >1000-fold selectivity versus an insulin variant lacking the N-terminal phenylalanine (Phe) residue. The crystal structure of the …
Determining Protease Substrate Selectivity And Inhibition By Label-Free Supramolecular Tandem Enzyme Assays, Garima Ghale, Vijayakumar Ramalingam, Adam R. Urbach, Werner M. Nau
Determining Protease Substrate Selectivity And Inhibition By Label-Free Supramolecular Tandem Enzyme Assays, Garima Ghale, Vijayakumar Ramalingam, Adam R. Urbach, Werner M. Nau
Chemistry Faculty Research
An analytical method has been developed for the continuous monitoring of protease activity on unlabeled peptides in real time by fluorescence spectroscopy. The assay is enabled by a reporter pair comprising the macrocycle cucurbit[7]uril (CB7) and the fluorescent dye acridine orange (AO). CB7 functions by selectively recognizing N-terminal phenylalanine residues as they are produced during the enzymatic cleavage of enkephalin-type peptides by the metalloendopeptidase thermolysin. The substrate peptides (e.g., Thr-Gly-Ala-Phe-Met-NH2) bind to CB7 with moderately high affinity (K ≈ 104 M–1), while their cleavage products (e.g., Phe-Met-NH2) bind very tightly (K …