Details

Autor(en) / Beteiligte

• Acharya, Bipul R
• Bhattacharyya, Bhabatarak
• Chakrabarti, Gopal

Titel

The Natural Naphthoquinone Plumbagin Exhibits Antiproliferative Activity and Disrupts the Microtubule Network through Tubulin Binding

Ist Teil von

• Biochemistry (Easton), 2008-07, Vol.47 (30), p.7838-7845

Ort / Verlag

United States: American Chemical Society

Erscheinungsjahr

2008

Quelle

MEDLINE

Beschreibungen/Notizen

• Plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone), a naphthoquinone isolated from the roots of Plumbaginaceae plants, has potential antiproliferative activity against several tumor types. We have examined the effects of plumbagin on cellular microtubules ex vivo as well as its binding with purified tubulin and microtubules in vitro. Cell viability experiments using human non-small lung epithelium carcinoma cells (A549) indicated that the IC50 value for plumbagin is 14.6 μM. Immunofluorescence studies using an antitubulin FITC conjugated antibody showed a significant perturbation of the interphase microtubule network in a dose dependent manner. In vitro polymerization of purified tubulin into microtubules is inhibited by plumbagin with an IC50 value of 38 ± 0.5 μM. Its binding to tubulin quenches protein tryptophan fluorescence in a time and concentration dependent manner. Binding of plumbagin to tubulin is slow, taking 60 min for equilibration at 25 °C. The association reaction kinetics is biphasic in nature, and the association rate constants for fast and slow phases are 235.12 ± 36 M−1 s−1 and 11.63 ± 11 M−1 s−1 at 25 °C respectively. The stoichiometry of plumbagin binding to tubulin is 1:1 (mole:mole) with a dissociation constant of 0.936 ± 0.71 μM at 25 °C. Plumbagin competes for the colchicine binding site with a K i of 7.5 μM as determined from a modified Dixon plot. Based on these data we conclude that plumbagin recognizes the colchicine binding site to tubulin. Further study is necessary to locate the pharmacophoric point of attachment of the inhibitor to the colchicine binding site of tubulin.