Putative binding mode of various ligands in AT₁ receptor.

<p>Transmenbrane (TM)1–7 are shown as colored ribbons: blue (TM1), cyan (TM2), green (TM3), lime green (TM4), yellow (TM5), orange (TM6) and red (TM7). Sidechains of the focused residues are shown as gray lines, and ligands are represented as magenta sticks. The hydrogen bonds and electrostatic interactions are indicated by yellow dotted lines. The distance between Leu¹¹² and the hydroxyphenyl moiety of R794847 is represented as a green dotted line. The unit of distance was Å. (a) without ligand, (b) olmesartan, (c) R239470, (d) R794847.</p

Binding affinities (<i>K</i>_d) of Ang II, olmesartan, R239470 and R794847 to AT₁ wild-type (WT) and mutant receptors.

<p>Binding affinities (<i>K</i>_d) of Ang II, olmesartan, R239470 and R794847 to AT₁ wild-type (WT) and mutant receptors.</p

Binding affinities (<i>K</i>_d) of Ang II and R compounds to AT₁ wild-type (WT) and mutant receptors.

<p>Binding affinities (<i>K</i>_d) of Ang II and R compounds to AT₁ wild-type (WT) and mutant receptors.</p

Binding affinities (<i>K</i>_d) of Ang II, olmesartan, R239470 and R794847 to AT₁ wild-type (WT) and mutant receptors.

<p>Binding affinities (<i>K</i>_d) of Ang II, olmesartan, R239470 and R794847 to AT₁ wild-type (WT) and mutant receptors.</p

Construction of olmesartan-related compounds.

<p>Construction of olmesartan-related compounds.</p

% inositol phosphate (IP) production with or without 1 µM of olmesartan and olmesartan-related compounds in COS1 cells transiently expressing the wild-type (WT) and N111G AT₁ receptor.

<p>The test compounds were added 1 h before the measurement of IP. 100% IP production indicates basal IP production in WT (1,033±149 cpm) and N111G (2,087±414 cpm) AT₁ receptor-transfected cells. Ang II (0.1 µM)-induced maximum IP production in WT and N111G AT₁ receptor-transfected cells were 4,037±479 cpm and 4,138±362 cpm, respectively. n=4–7, *p<0.05 vs. no treatment.</p

Changes in cysteine accessibility in COS1 cells expressing C76A/C289A mutant receptors in which the transmenbrane 3 residues from Ala¹⁰⁶ to Phe¹¹⁷, except for position 113, were successively replaced by cysteine.

<p>1 µM of ARBs, n=4–9, *p<0.05 vs. no treatment. #p<0.05 vs. C76A/C289A.</p

A. Summary of cysteine accessibilityat Ser¹⁰⁹, Leu¹¹², Val¹¹⁶ and Phe¹¹⁷ with olmesartan, R239470 and R794847 compared to ligand (-).

<p>Less and More indicate that the AT₁ receptor is less or more accessible to the ligand pocket, respectively. B. Helical wheel representation of transmenbrane (TM)3 cysteine residues for ligand (-), olmesartan, R239470 and R794847. Olmesartan induced the outward displacement and counterclockwise rotation of TM3. Dotted circle indicates the original position of TM3 without ligand. Accessible area indicates a site of ligand pocket. C. Schematic view of TM3 movement derived from the SCAM results. TM3 without ligand is shown as a gray ribbon and TM3 with various ligands is represented as green ribbons. The ligands are colored magenta. (a) without ligand, (b) olmesartan, (c) R239470, (d) R794847.</p