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Selective chromogenic and fluorogenic peptide substrates for the assay of cysteine peptidases in complex mixtures
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SYSNO ASEP 0424813 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Selective chromogenic and fluorogenic peptide substrates for the assay of cysteine peptidases in complex mixtures Author(s) Semashko, T. A. (RU)
Vorotnikova, E. A. (RU)
Sharikova, V. F. (RU)
Vinokurov, Konstantin (BC-A) RID, ORCID
Smirnova, Y. A. (RU)
Dunaevsky, Y. E. (RU)
Belozersky, M. A. (RU)
Oppert, B. (US)
Elpidina, E. N. (RU)
Filippova, I. Y. (RU)Number of authors 10 Source Title Analytical Biochemistry. - : Elsevier - ISSN 0003-2697
Roč. 449, č. 1 (2014), s. 179-187Number of pages 9 s. Language eng - English Country US - United States Keywords cysteine peptidases ; substrates of peptidases ; selective peptide substrates Subject RIV CE - Biochemistry Institutional support BC-A - RVO:60077344 UT WOS 000332816100026 EID SCOPUS 84893337071 DOI 10.1016/j.ab.2013.12.032 Annotation This study describes the design, synthesis, and use of selective peptide substrates for cysteine peptidases of the C1 papain family, important in many biological processes. The structure of the newly synthesized substrates is Glp-Xaa-Ala-Y (where Glp = pyroglutamyl; Xaa = Phe or Val; and Y = pNA [p -nitroanilide], AMC [4-amino-7-methylcoumaride], or AFC [4-amino-7- trifluoromethyl-coumaride]). Substrates were synthesized enzymatically to guarantee selectivity of the reaction and optical purity of the target compounds, simplifying the scheme of synthesis and isolation of products. The hydrolysis of the synthesized substrates was evaluated by C1 cysteine peptidases from different organisms and with different functions, including plant enzymes papain, bromelain, ficin, and mammalian lysosomal cathepsins B and L. The new substrates were selective for C1 cysteine peptidases and were not hydrolyzed by serine, aspartic, or metallo peptidases. We demonstrated an application of the selectivity of the synthesized substrates. Workplace Biology Centre (since 2006) Contact Dana Hypšová, eje@eje.cz, Tel.: 387 775 214 Year of Publishing 2015 Electronic address http://www.sciencedirect.com/science/article/pii/S0003269713006180#
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