Characterization of cysteine proteases in Malian medicinal plants

• Published in: Journal of ethnopharmacology Vol. 107; no. 2; pp. 189 - 198
• Main Authors: Bah, Sékou, Paulsen, Berit S., Diallo, Drissa, Johansen, Harald T.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Ireland Ltd 19-09-2006; Elsevier
• Subjects: anthelmintics; Biological and medical sciences; Cissus; Cissus quadrangularis; Cysteine Endopeptidases - isolation & purification; Cysteine Endopeptidases - metabolism; Cysteine Endopeptidases - pharmacology; Cysteine proteases; Cysteine Proteinase Inhibitors - pharmacology; cysteine proteinases; Electrophoresis, Polyacrylamide Gel; Entada; Entada africana; Enzyme Activation; enzyme inhibitors; ethnobotany; General pharmacology; herbal medicines; Mali; Malian medicinal plants; Medical sciences; medicinal plants; medicinal properties; Peptides - chemistry; Peptides - metabolism; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; plant extracts; Plant Extracts - isolation & purification; Plant Extracts - metabolism; Plant Extracts - pharmacology; Plants, Medicinal - chemistry; Plants, Medicinal - classification; Plants, Medicinal - enzymology; Protease inhibitors; schistosomiasis; Schistosomicides - isolation & purification; Schistosomicides - pharmacology; Securidaca; Securidaca longepedunculata; Stylosanthes; Stylosanthes erecta; Substrate Specificity; Synthetic substrates; traditional medicine; Mali; Cysteine proteases; Synthetic substrates; Malian medicinal plants; Protease inhibitors; Thiol; Cysteine; Enzyme; Pharmacognosy; Medicinal plant; Sulfur containing aminoacid; Peptidases; Plant origin; Hydrolases; Protease inhibitor
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2006.03.008

Extracts form 10 different Malian medicinal plants with a traditional use against schistosomiasis were investigated for their possible content of proteolytic activity. The proteolytic activity was studied by measuring the hydrolysis of two synthetic peptide substrates Z–Ala–Ala–Asn–NHMec and Z–Phe–Arg–NHMec. Legumain- and papain-like activities were found in all tested crude extracts except those from
Entada africana, with the papain-like activity being the strongest.
Cissus quadrangularis,
Securidaca longepedunculata and
Stylosanthes erecta extracts showed high proteolytic activities towards both substrates. After gel filtration the proteolytic activity towards the substrate Z–Ala–Ala–Asn–NHMec in root extract of
Securidaca longepedunculata appeared to have Mr of 30 and 97
kDa, while the activity in extracts from
Cissus quadrangularis was at 39
kDa. Enzymatic activity cleaving the substrate Z–Phe–Arg–NHMec showed apparent Mr of 97 and 26
kDa in extracts from roots and leaves of
Securidaca longepedunculata, while in
Cissus quadrangularis extracts the activity eluted at 39 and 20
kDa, with the highest activity in the latter. All Z–Phe–Arg–NHMec activities were inhibited by E-64 but unaffected by PMSF. The legumain activity was unaffected by E-64 and PMSF. The SDS-PAGE analysis exhibited five distinct gelatinolytic bands for
Cissus quadrangularis extracts (115, 59, 31, 22 and 20
kDa), while two bands (59 and 30
kDa) were detected in
Securidaca longepedunculata extracts. The inhibition profile of the gelatinolytic bands and that of the hydrolysis of the synthetic substrates indicate the cysteine protease class of the proteolytic activities. Several cysteine protease activities with different molecular weights along with a strong variability of these activities between species as well as between plant parts from the same species were observed.