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Volumn 14, Issue 20, 2004, Pages 5113-5116

Inhibition of lysosomal cysteine proteases by chrysotherapeutic compounds: A possible mechanism for the antiarthritic activity of Au(I)

Author keywords

[No Author keywords available]

Indexed keywords

AURANOFIN; CYSTEINE PROTEINASE; GOLD COMPLEX; LYSOSOME ENZYME; THIOMALIC ACID;

EID: 4544310043     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2004.07.073     Document Type: Article
Times cited : (61)

References (32)
  • 26
    • 4544373050 scopus 로고    scopus 로고
    • note
    • Enzyme activity assays: Standard conditions used to determine the activity of the purified enzymes against Z-Phe-Arg-MCA are described below. Substrate stocks were dissolved in DMSO and aliquots taken such that each reaction had the same amount of DMSO, which never exceeded 5% of the total reaction volume. The buffer used for all studies contained 20 mM sodium acetate, pH 5.5, 50 mM sodium chloride, 1.0 mM EDTA, and 0.01% Brij 35, with 400 μM DTT added to the buffer just prior to each reaction. In each reaction, the concentration of enzyme was fixed between 1 and 10 nM and the substrate concentration varied from 5 nM to 100 μM. The increase in fluorescence as the substrate was hydrolyzed was measured every 60 s for 30 min. Fluorescence was converted into concentration of 7-amino-4-methylcoumarin (MCA) produced using a standard curve of MCA fluorescence, and the data was fit to the Michaelis-Menten equation, providing kinetic parameters for each enzyme
  • 27
    • 4544383477 scopus 로고    scopus 로고
    • note
    • 2Cl, which was insoluble at concentrations above 1 μM
  • 30
    • 4544291801 scopus 로고    scopus 로고
    • note
    • The possibility of reversing the Au(I) inhibition of Paramecium tetraurelia cathepsin L was investigated by first preincubating the enzyme with 50 μM myochrysin for 1 h, then adding between 20 nM and 200 μM of thiomalic acid or reduced glutathione and incubating another 1 h. Finally, substrate was added to each mixture and the rate of reaction was determined
  • 31
    • 4544330300 scopus 로고    scopus 로고
    • note
    • To determine the mode of inhibition, four different reactions were run where the substrate concentration was varied from 20 to 90 μM at inhibitor concentrations of 300 nM, 3, 30, and 300 μM


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.