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Fig 1.

The chemical structure of nine bufadienolides.

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Fig 1 Expand

Fig 2.

Quenching effect of bufadienolides against human serum albumin (HSA) fluorescence.

Experiments were conducted at 298 K. λex = 280 nm; HSA, 2.0 × 10–6 M; a–g = 0, 10, 20, 40, 80, 160 and 200 × 10–6 M for gamabufotalin (A), arenobufagin (B), bufotalin (C), cinobufagin (D), hellebrigenin (E), telocinobufagin (F), resibufogenin (G) and bufalin (H); and a–g = 0, 6.3, 12.5, 25, 50 and 200×10–6 M for desacety-bufotalin (I).

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Fig 2 Expand

Table 1.

Stern-Volmer quenching constants (KSV) of the interaction of bufadienolides with HSA at different temperatures.

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Table 1 Expand

Fig 3.

Relationship between bufadienolide binding affinities and the number of binding sites for human serum albumin (HSA).

logKa = binding affinity; Ka = binding constant; n = number of binding sites; and R = correlation coefficient.

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Fig 3 Expand

Table 2.

The binding constants of bufadienolides on HSA fluorescence.

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Table 2 Expand

Table 3.

Thermodynamic Parameters between bufadienolides-HSA interaction at pH 7.4.

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Table 3 Expand

Table 4.

Structural parameters and binding constants of bufadienolides for HSA at 298 K.

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Table 4 Expand

Fig 4.

Relationship between hydrogen (H-) bond acceptor/donor numbers and bufadienolide binding affinities for human serum albumin (HSA).

The H-bond acceptor/donor numbers were taken from the PubChem Public Chemical Database. logKa = binding affinity; Ka = binding constant; and R = correlation coefficient.

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Fig 4 Expand

Fig 5.

Bufadienolide docking to site I in human serum albumin (HSA).

The detailed binding conformations are shown for (A) gamabufotalin, (B) bufalin, (C) bufotalin, (D) resibufogenin and (E) cinobufagin. In each case, the drug is shown as a stick model. Key interaction amino acid residues (with names) are shown as sticks that are color-coded by atom type, and green dashed lines indicate hydrogen bonds (H-bonds).

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Fig 5 Expand

Table 5.

Molecular docking of various bufadienolides to HSA.

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Table 5 Expand