Design and Application of a Novel Chaicone Derivative Fluorescent Probe for Aminopeptidase N
Aminopeptidase N(APN)is a widely expressed hydrolytic enzyme in organisms that can hydrolyze neutral or alkaline amino acids at the N-terminus of protein polypeptide chains.Overexpressed APN may indicate disorders of biosystems,such as various tumors,drug-induced liver injury(DILI),and glomerulonephritis,and is widely used for early diagnosis and therapy of related diseases.Particularly,elevated APN level in urine is becoming one of the key markers of early kidney injury due to its special sensitivity to glomerulonephritis than other proteins.Herein,based on the structure of 4-amino chalcone,a novel fluorescent probe Ala-OMN for detecting APN was designed and synthesized by caging the 4-amino of OMN by alanine.Ala-OMN is non-fluorescent in phosphate buffered saline(PBS).After reaction with APN,the caging group alanine was hydrolyzed and OMN was released,which further combined with bovine serum protein(BSA)to form a1∶1 complex,concomitantly with a bright green fluorescence signal for indicating APN.Ala-OMN exhibits excellent sensitivity,selectivity,and anti-interference ability in response to APN,with a detection limit as low as 0.058 ng/mL.Ala-OMN has been successfully applied to detect APN concentration in urine samples,and the results are in good agreement with commercial reagent kits,suggesting that Ala-OMN could be a new diagnostic reagent for the clinical detection of early glomerulonephritis.
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