A dityrosine-based substrate for a protease assay

Application for the selective assessment of papain and chymopapain activity

Chan Jin Kim, Dong Ik Lee, Chang-Ha Lee, Ik Sung Ahn

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

N,N'-diBoc-dityrosine (DBDY), which was synthesized by the oxidative C-C coupling of 2 N-Boc-l-tyrosine molecules, was conjugated with two isoniazid (INH) molecules. Due to the quenching effect of INH, DBDY-(INH) 2 lacks the fluorescence of DBDY. As such, it was tested for use in the detection of proteases by measuring fluorescence recovery. In this study, serine proteases (chymotrypsin, trypsin, subtilisin, and proteinase K), metalloproteases (thermolysin and carboxypeptidase A, dispase, and collagenase), aspartic proteases (pepsin and aspergillopepsin) and cysteine proteases (papain and chymopapain) were chosen. Reported optimum assay conditions were chosen for each enzyme. Only papain and chymopapain catalyzed the hydrolysis of DBDY-(INH) 2 and led to fluorescence recovery, possibly due to their extensive binding sites and the INH-mediated inhibition of metalloproteases and aspartic proteases.

Original languageEnglish
Pages (from-to)101-107
Number of pages7
JournalAnalytica Chimica Acta
Volume723
DOIs
Publication statusPublished - 2012 Apr 20

Fingerprint

Chymopapain
Papain
Assays
Peptide Hydrolases
fluorescence
assay
substrate
Fluorescence
Metalloproteases
Substrates
Carboxypeptidases A
Thermolysin
Subtilisin
Recovery
Endopeptidase K
Molecules
Cysteine Proteases
Pepsin A
hydrolysis
Isoniazid

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

Cite this

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title = "A dityrosine-based substrate for a protease assay: Application for the selective assessment of papain and chymopapain activity",
abstract = "N,N'-diBoc-dityrosine (DBDY), which was synthesized by the oxidative C-C coupling of 2 N-Boc-l-tyrosine molecules, was conjugated with two isoniazid (INH) molecules. Due to the quenching effect of INH, DBDY-(INH) 2 lacks the fluorescence of DBDY. As such, it was tested for use in the detection of proteases by measuring fluorescence recovery. In this study, serine proteases (chymotrypsin, trypsin, subtilisin, and proteinase K), metalloproteases (thermolysin and carboxypeptidase A, dispase, and collagenase), aspartic proteases (pepsin and aspergillopepsin) and cysteine proteases (papain and chymopapain) were chosen. Reported optimum assay conditions were chosen for each enzyme. Only papain and chymopapain catalyzed the hydrolysis of DBDY-(INH) 2 and led to fluorescence recovery, possibly due to their extensive binding sites and the INH-mediated inhibition of metalloproteases and aspartic proteases.",
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A dityrosine-based substrate for a protease assay : Application for the selective assessment of papain and chymopapain activity. / Kim, Chan Jin; Lee, Dong Ik; Lee, Chang-Ha; Ahn, Ik Sung.

In: Analytica Chimica Acta, Vol. 723, 20.04.2012, p. 101-107.

Research output: Contribution to journalArticle

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