Figure 4Results of Glide XP docking of colchicine, noscapine, and Br-noscapine into the colchicine binding site of tubulin

2D representations of the binding site with amino acid residues involved in the interaction of (A) colchicine and (B) Br-noscapine reveal identical sets of amino acids. (C) 3D representations of the binding site amino acids within a 4Å distances from superimposed conformations of docked noscapine (blue) and Br-noscapine (green). The co-crystal structure of colchicine is represented in red color. Some of the amino acids (Leu248, Cys241, Val318, Ala316 and Val238) are not shown for better clarity. (D) Overlapped docking poses of colchicine, noscapine, and Br-noscapine obtained form Glide docking onto co-crystal of colchicine within the colchicine binding site of tubulin. Tubulin is represented as Macromodel surface according to residue charge (electropositive charge, blue; neutral, yellow) as implemented in Maestro.

Molecular modeling and competition binding study of Br-noscapine and colchicine provides insight into noscapinoid-tubulin binding site

J Mol Graph Model. 2011 June;29(7):947-955.
Fig. 3.

Interaction of colchicine with the human 5-HT_3AR. A, colchicine enhances currents evoked by 0.5 μM 5-HT in oocytes expressing the human 5-HT_3AR. Serotonin (0.5 μM) was applied for 30 s to obtain a baseline current. Colchicine (300 μM) was coapplied with 5-HT (0.5 μM) for 30 s. A potentiation of ∼75% of the baseline current was observed in this receptor. Washout of colchicine-induced potentiation was observed with the next application of 5-HT 5 min later. B, concentration-response curve for colchicine (10–1000 μM) was generated in oocytes expressing human 5-HT_3ARs (n = 4–5). An EC₅₀ for colchicine of 226.6 ± 25.1 μM and a Hill coefficient of 2.38 ± 0.64 were determined. C, potentiating actions of colchicine at the human 5-HT_3AR is 5-HT concentration dependent and microtubule independent. Colchicine (300 μM) was coapplied with 5-HT (0.5, 1, 3, or 10 μM) for 30 s (n = 3–4). The effect of colchicine was expressed as a percentage of the baseline current produced by the respective concentration of 5-HT alone.

Colchicine: A Novel Positive Allosteric Modulator of the Human 5-Hydroxytryptamine3A Receptor

J Pharmacol Exp Ther. 2009 May;329(2):838-847.
FIGURE 5

(A) Effect of BbTb (- - -) and CeTb (·····) on fluorescence of colchicine. Tubulin (1 μM) was added to colchicine (150 μM) in PME buffer in presence of 0.1 mM GTP at 25°C and enhancement in colchicine fluorescence at 440 nm was monitored as function of time. (B) Effect of colchicine on intrinsic fluorescence of BbTb (- - -) and CeTb (·····). Colchicine (150 μM) was added to tubulin (1 μM) in PME buffer in presence of 0.1 mM GTP at 25°C and the quenching of intrinsic protein fluorescence at 335 nm was monitored as function of time.

Characterization of the Colchicine Binding Site on Avian Tubulin Isotype ?VI

Biochemistry. 2010 April 6;49(13):2932-2942.
Fig. 4

Localization of man II and ALP in McA-RH 7777 cells incubated in basal medium after 4-hr colchicine treatment. (a) Localization of man II in McA-RH 7777 cells incubated for 8 hr in basal medium after 4-hr colchicine treatment. Man II is localized in numerous small dots scattered throughout the cytoplasm in most cells, although man II-containing dots aggregate around the nucleus in a few cells. (b) Localization of ALP in McA-RH 7777 cells incubated for 8 hr in basal medium after 4-hr colchicine treatment. Intense staining for ALP is recognized at intercellular contacts and small dots containing ALP is observed in the cytoplasm. In some cells, positive reactions for ALP are also observed in areas of the plasma membrane not in contact with other cells. (c) Merged image of (a) and (b). (d) Localization of man II in McA-RH 7777 cells incubated for 20 hr in basal medium after 4-hr colchicine treatment. Small dots containing man II are still scattered throughout the cytoplasm in other cells, although they aggregate in the perinuclear cytoplasm in some cells. (e) Localization of ALP in McA-RH 7777 cells incubated for 20 hr in basal medium after 4-hr colchicine treatment. Large spots with intense staining for ALP reappear in the perinuclear cytoplasm in numerous cells. (f) Merged image of (d) and (e). (g) Localization of man II in McA-RH 7777 cells incubated for 26 hr in basal medium after 4-hr colchicine treatment. (h) Localization of ALP in McA-RH 7777 cells incubated for 26 hr in basal medium after 4-hr colchicine treatment. Localization patterns of man II and ALP are similar to those in McA-RH 7777 cells cultured only in basal medium. (i) Merged image of (g) and (h). Bar=50 µm.

Effects of Colchicine on Localization of Alkaline Phosphatase in McA-RH 7777 Rat Hepatoma Cells

Acta Histochem Cytochem. 2008 December 27;41(6):149-155.

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