Study of a novel antiosteoporosis screening model targeted on cathepsin K / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 273-280, 2004.
Article
Dans Anglais
| WPRIM
| ID: wpr-329635
ABSTRACT
<p><b>OBJECTIVE</b>To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs.</p><p><b>METHODS</b>To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing. Cathepsin K was expressed in a high amount in E. coli after IPTG induction, and was purified to near homogenetity through resolution and column purification. The specificity of the protein was shown by Western blotting experiment. The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain.</p><p><b>RESULTS</b>With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects. The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K.</p><p><b>CONCLUSIONS</b>The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Ostéoporose
/
Dosage biologique
/
Papaïne
/
Cathepsines
/
Expression des gènes
/
Lignée cellulaire
/
Technique de Western
/
Clonage moléculaire
/
Similitude de séquences d'acides aminés
/
Analyse de séquence d'ADN
Type d'étude:
Etude diagnostique
/
Étude de dépistage
Limites du sujet:
Humains
langue:
Anglais
Texte intégral:
Biomedical and Environmental Sciences
Année:
2004
Type:
Article
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