A new, robust and reliable methodology for three-protease screening in a single-enzyme mode has been developed and verified, employing a multi-purpose peptide probe with three selectively cleavable sites furnished with four fluorophores. A triple-FRET-based single-excitation quadruple-emission concept for unambiguous sensing of trypsin, chymotrypsin and caspase-8 in the lowest detectable concentrations of 0.5 ng mL-1, 0.2 μg mL-1, and 2 U mL-1, respectively, has been applied and graphically depicted. Then the developed 4-dye probe has been also studied from the perspective of simultaneous two-protease screening, which was found only partially feasible, primarily due to unselective chymotrypsin cleavage.

Department of Organic Chemistry Faculty of Science Palacký University Olomouc 17 Listopadu 12 771 46 Olomouc Czech Republic

Zobrazit více v PubMed

López-Otín C. Overall C. M. Nat. Rev. Mol. Cell Biol. 2002;3:509–519. PubMed

Turk B. Nat. Rev. Drug Discovery. 2006;5:785–799. PubMed

Vasiljeva O. Reinheckel T. Peters C. Turk D. Turk V. Turk B. Curr. Pharm. Des. 2007;13:387–403. PubMed

Carmona A. K. Juliano M. A. Juliano L. An. Acad. Bras. Cienc. 2009;81:381–392. PubMed

Clegg R. M., in Reviews in Fluorescence 2006, ed. C. D. Geddes and J. R. Lakowicz, Springer, Boston, MA, 2006, pp. 1–45

Wu L. Huang C. Emery B. P. Sedgwick A. C. Bull S. D. He X. P. Tian H. Yoon J. Sessler J. L. James T. D. Chem. Soc. Rev. 2020;49:5110–5139. PubMed PMC

Lerner E. Barth A. Hendrix J. Ambrose B. Birkedal V. Blanchard S. C. Börner R. Chung H. S. Cordes T. Craggs T. D. Deniz A. A. Diao J. Fei J. Gonzalez R. L. Gopich I. V. Ha T. Hanke C. A. Haran G. Hatzakis N. S. Hohng S. Hong S. C. Hugel T. Ingargiola A. Joo C. Kapanidis A. N. Kim H. D. Laurence T. Lee N. K. Lee T. H. Lemke E. A. Margeat E. Michaelis J. Michalet X. Myong S. Nettels D. Peulen T. O. Ploetz E. Razvag Y. Robb N. C. Schuler B. Soleimaninejad H. Tang C. Vafabakhsh R. Lamb D. C. Seidel C. A. M. Weiss S. Boudker O. Elife. 2021;10:1–69. PubMed PMC

Liu L. He F. Yu Y. Wang Y. Front. Bioeng. Biotechnol. 2020;8:1–17. PubMed PMC

Imani M. Mohajeri N. Rastegar M. Zarghami N. Anal. Biochem. 2021;630:114323. PubMed

Li S. Y. Liu L. H. Cheng H. Li B. Qiu W. X. Zhang X. Z. Chem. Commun. 2015;51:14520–14523. PubMed

Cheng H. Li S. Y. Zheng H. R. Li C. X. Xie B. R. Chen K. W. Li B. Zhang X. Z. Anal. Chem. 2017;89:4349–4354. PubMed

Okorochenkova Y. Porubský M. Benická S. Hlaváč J. Chem. Commun. 2018;54:7589–7592. PubMed

Megia-Fernandez A. Mills B. Michels C. Chankeshwara S. V. Krstajić N. Haslett C. Dhaliwal K. Bradley M. Org. Biomol. Chem. 2018;16:8056–8063. PubMed PMC

Jokerst J. Cheng Y. Borum R. M. Clark A. E. Jin Z. Moore C. Fajtová P. O'Donoghue A. J. Carlin A. F. Angew. Chem., Int. Ed. 2022;61(1–8):e202113617. PubMed PMC

Algar W. R. Krause K. D. Annu. Rev. Anal. Chem. 2022;15:17–36. PubMed

Wu M. Massey M. Petryayeva E. Algar W. R. J. Phys. Chem. C. 2015;119:26183–26195.

Algar W. R. Wegner D. Huston A. L. Blanco-Canosa J. B. Stewart M. H. Armstrong A. Dawson P. E. Hildebrandt N. Medintz I. L. J. Am. Chem. Soc. 2012;134:1876–1891. PubMed

Wu M. Petryayeva E. Algar W. R. Anal. Chem. 2014;86:11181–11188. PubMed

Algar W. R. Ancona M. G. Malanoski A. P. Susumu K. Medintz I. L. ACS Nano. 2012;6:11044–11058. PubMed

Massey M. Kim H. Conroy E. M. Algar W. R. J. Phys. Chem. C. 2017;121:13345–13356.

Tsai H. Y. Kim H. Massey M. Krause K. D. Algar W. R. Methods Appl. Fluoresc. 2019;7(1–26):042001. PubMed

Wu M. Algar W. R. Anal. Chem. 2015;87:8078–8083. PubMed

He G. Guo D. He C. Zhang X. Zhao X. Duan C. Angew. Chem., Int. Ed. 2009;48:6132–6135. PubMed

Gong Y. J. Zhang X. B. Mao G. J. Su L. Meng H. M. Tan W. Feng S. Zhang G. Chem. Sci. 2016;7:2275–2285. PubMed PMC

Sánchez-Mundo M. L. Bautista-Muñoz C. Jaramillo-Flores M. E. CyTA--J. Food. 2015;13:578–587.

Atrooz O. M. Alomari F. N. J. Appl. Biol. Biotechnol. 2020;8:33–37.

Christopeit T. Øverb K. Danielson U. H. Nilsen I. W. Mar. Drugs. 2013;11:4279–4293. PubMed PMC

Milićević D. Hlaváč J. Chemosensors. 2021;9(1–13):119.

Milićević D. Hlaváč J. Chemosensors. 2021;9(1–11):359.

A new multi-purpose FRET fluorescent probe for the simultaneous detection of proteases

Novel Peptide-Based Fluorescent Probe for Simultaneous Sensing of Chymotrypsin and Hydrogen Peroxide