Journal: The Journal of Biological Chemistry

Article Title: Mapping the binding site of the P2X receptor antagonist PPADS reveals the importance of orthosteric site charge and the cysteine-rich head region

doi: 10.1074/jbc.RA118.003737

Figure Lengend Snippet: Comparison of proposed PPADS and TNP-ATP binding modes. a , proposed PPADS binding mode for P2X1R. b , for comparison the X-ray structure of chicken P2X7 receptor with TNP-ATP bound is shown. Equivalent lysine residues involved in salt bridges to sulfonate groups of PPADS ( a ) and phosphate groups of TNP-ATP are labeled. c , visualization of electrostatic potential for the P2X1R model used in a . The electrostatic potential was calculated using the Protein Data Bank 2PQR . d, as in c , but PPADS was omitted.

Article Snippet: Finally, the total and biotinylated P2X1R proteins in apyrase and PPADS treatment were quantified by Western blotting with an anti-P2X1R antibody (1:2000) (APR-001 lot number AN0802, Alomone, Jerusalem, Israel) as described previously ( ).

Techniques: Binding Assay, Labeling

Journal: The Journal of Biological Chemistry

Article Title: Mapping the binding site of the P2X receptor antagonist PPADS reveals the importance of orthosteric site charge and the cysteine-rich head region

doi: 10.1074/jbc.RA118.003737

Figure Lengend Snippet: Comparison of proposed PPADS and TNP-ATP binding modes. a , proposed PPADS binding mode for P2X1R. b , for comparison the X-ray structure of chicken P2X7 receptor with TNP-ATP bound is shown. Equivalent lysine residues involved in salt bridges to sulfonate groups of PPADS ( a ) and phosphate groups of TNP-ATP are labeled. c , visualization of electrostatic potential for the P2X1R model used in a . The electrostatic potential was calculated using the Protein Data Bank 2PQR . d, as in c , but PPADS was omitted.

Article Snippet: Finally, the total and biotinylated P2X1R proteins in apyrase and PPADS treatment were quantified by Western blotting with an anti-P2X1R antibody (1:2000) (APR-001 lot number AN0802, Alomone, Jerusalem, Israel) as described previously ( ).

Techniques: Binding Assay, Labeling