title: Kinetic and structural analysis of fluorescent peptides on cotton cellulose nanocrystals as elastase sensors.
abstract: Human neutrophil elastase (HNE) and porcine pancreatic elastase (PPE) are serine proteases with destructive proteolytic activity. Because of this activity, there is considerable interest in elastase sensors. Herein we report the synthesis, characterization, and kinetic profiles of tri- and tetrapeptide substrates of elastase as glycine-esterified fluorescent analogs of cotton cellulose nanocrystals (CCN). The degree of substitution of peptide incorporated in CCN was 3-4 peptides per 100 anhydroglucose units. Glycine and peptide-cellulose-nanocrystals revealed crystallinity indices of 79 and 76%, respectively, and a crystallite size of 58.5 Å. A crystallite model of the peptide-cellulose conjugate is shown. The tripeptide conjugate of CCN demonstrated five-fold greater efficiency in HNE than the tripeptide in solution judged by its kcat/Km of 33,515. The sensor limits of detection at 2mg of the tri- and tetrapeptide CCN conjugates over a 10 min reaction time course were 0.03 U/mL PPE and 0.05 U/mL HNE, respectively.
authors.affiliation: USDA-ARS, Southern Regional Research Center, Cotton Chemistry & Utilization, 1100 Robert E. Lee Blvd., New Orleans, LA 70124, United States. Electronic address: vince.edwards@ars.usda.gov.
authors.forename: J Vincent
authors.initials: JV
authors.lastname: Edwards
authors.affiliation: USDA-ARS, Southern Regional Research Center, Cotton Chemistry & Utilization, 1100 Robert E. Lee Blvd., New Orleans, LA 70124, United States.
authors.forename: Nicolette T
authors.initials: NT
authors.lastname: Prevost
authors.affiliation: USDA-ARS, Southern Regional Research Center, Cotton Chemistry & Utilization, 1100 Robert E. Lee Blvd., New Orleans, LA 70124, United States.
authors.forename: Alfred D
authors.initials: AD
authors.lastname: French
authors.affiliation: USDA-ARS, Southern Regional Research Center, Cotton Chemistry & Utilization, 1100 Robert E. Lee Blvd., New Orleans, LA 70124, United States.
authors.forename: Monica
authors.initials: M
authors.lastname: Concha
authors.affiliation: USDA-ARS, Southern Regional Research Center, Cotton Chemistry & Utilization, 1100 Robert E. Lee Blvd., New Orleans, LA 70124, United States.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Condon
mesh_terms: D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D002482:Cellulose / Q000737:chemistry*; D032561:Cotton Fiber; D005453:Fluorescence; D006801:Humans; D007700:Kinetics; D019272:Leukocyte Elastase / Q000032:analysis* / Q000737:chemistry; D008958:Models, Molecular; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*
chemical_list: D015415:Biomarkers; D010455:Peptides; D002482:Cellulose; D019272:Leukocyte Elastase
keywords: Biosensor; Cotton/cellulose; Human neutrophil elastase; Nanocrystals