(A) Chemical structures of uridine diphosphate (UDP) and UDP-glucose (UDP-Glc). (B) Concentration-response curves of calcium mobilization for UDP or UDP-Glc in HEK293 cells transiently …
Potency of uridine diphosphate (UDP) or UDP-glucose (UDP-Glc) in HEK293 cells expressing P2Y purinoceptor 14 (P2Y14).
Potency of uridine diphosphate (UDP) or UDP-glucose (UDP-Glc) in HEK293 cells expressing P2Y purinoceptor 14 (P2Y14) wild-type (WT) and mutants.
Comparison of EC50s for uridine diphosphate-glucose (UDP-Glc) and UDP in HEK293 cells expressing P2Y purinoceptor 14 (P2Y14) mutants.
(A, B) Docking models of UDP (A) and UDP-Glc (B) to P2Y14 showing binding sites for UDP moieties. Protein is shown in cartoon representation. UDP (blue), UDP-Glc (salmon), and key residues (yellow) …
(A–B) Side (A) and top (B) views of representative models of apo P2Y14 and UDP-Glc-bound P2Y14. Receptor is shown as cartoon. Ligand and key residues are shown as sticks. Movement of the …
Potency of uridine diphosphate-glucose (UDP-Glc) in HEK293 expressing P2Y purinoceptor 14 (P2Y14) wild-type (WT) and mutants.
(A–B) Side (A) and top (B) views of representative models of apo P2Y14 and UDP-Glc-bound P2Y14. Receptor is shown as cartoon. Ligand and key residues are shown as sticks. Movement of the …
(A) Chemical structures of uridine diphosphate-glucose (UDP-Glc), UDP-glucuronic acid (GlcA), UDP-N-acetylglucosamine (GlcNAc), and UDP-galactose (Gal). (B) Concentration-response curves of calcium …
Potency of uridine diphosphate-glucuronic acid (UDP-GlcA), UDP-N-acetylglucosamine (GlcNAc), and UDP-galactose (Gal) in HEK293 expressing P2Y purinoceptor 14 (P2Y14).
Potency of uridine diphosphate-glucuronic acid (UDP-GlcA), UDP-N-acetylglucosamine (GlcNAc), and UDP-galactose (Gal) in HEK293 expressing P2Y purinoceptor 14 (P2Y14) wild-type (WT) and mutants.
Error bars indicate standard deviations involving three independent molecular dynamics (MD) simulation trajectories.
Three replicated simulations were indicated by different colors.
Putative hydrogen bonds and salt bridges are displayed as red dashed lines.
(A–D) Distribution of distance between key residues (K277, E278, R253, and K77) and a group of UDP-glucose (Glc) (A), UDP-glucuronic acid (GlcA) (B), UDP-N-acetylglucosamine (GlcNAc) (C), or …
(A) Sequence log of the alignment between P2Y12 and P2Y14. Sequences of P2Y12 and P2Y14 involving 359 species were collected for making sequence alignments. See Supplementary file 3 and Figure …
Potency of adenosine diphosphate-glucose (ADP-Glc), ADP-glucuronic acid (GlcA), and ADP-mannose (Man) in HEK293 expressing P2Y purinoceptor 12 (P2Y12) wild-type (WT) and mutants.
The neighbor joining tree of P2Y purinoceptor 12 (P2Y12) showing evolutionary range of species.
The neighbor joining tree of P2Y purinoceptor 13 (P2Y13) showing evolutionary range of species.
The neighbor joining tree of P2Y purinoceptor 14 (P2Y14) showing evolutionary range of species.
The neighbor joining tree of GPR87 showing evolutionary range of species.
(A) Conservation of each residue on P2Y12. The height of a letter is proportional to the relative frequency of that residue at a particular site. Residues of K2.60-D2.64-K7.35-E7.36 (KDKE) …
(A) Location of F2777.32 in the docking model of adenosine diphosphate-glucose (ADP-Glc)-bound P2Y12. Receptor is shown as cartoon. Ligands and key residues are shown as sticks. Residues of K2.60-D2.…
The height of a letter is proportional to the relative frequency of that residue at a particular site. Residues of K2.60, D2.64, K7.35, and E7.36 sites are labeled with red arrows. N.S. indicates …
The height of a letter is proportional to the relative frequency of that residue at a particular site. Residues of K/R2.60, D2.64, K/E7.35, and E7.36 sites are labeled with red arrows. See Supplement…
The height of a letter is proportional to the relative frequency of that residue at a particular site. Residues of KDKE sugar-binding motif are labeled with red arrows. See Supplementary file 3 for …
The height of a letter is proportional to the relative frequency of that residue at a particular site. Residues of KDKE sugar-binding motif are labeled with red arrows. See Supplementary file 3 for …
The height of a letter is proportional to the relative frequency of that residue at a particular site. Residues of KDKE sugar-binding motif are labeled with red arrows. See Supplementary file 3 for …
The height of a letter is proportional to the relative frequency of that residue at a particular site. Residues of KDKE sugar-binding motif are labeled with red arrows. See Supplementary file 3 for …
Sequences of P2Y14 and P2Y12 involving 359 species were collected for making sequence alignments. See Supplementary file 3 for species repertoire information. The height of a letter is proportional …
(A) A representative simulation model showing UDP in the sub-pocket of P2Y14. Key residues are highlighted in yellow. Two sub-pockets for ligand binding are marked with circles. (B) Massive atom …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Gene (Homo sapiens) | P2Y14 | GenBank | NM_001081455.2 | |
Gene (Homo sapiens) | P2Y12 | GenBank | NM_022788.5 | Optimized |
Strain, strain background (Escherichia coli) | Trans5α Chemically Competent Cell | TransGen Biotech | Cat.No: CD201-01 | |
Cell line (Homo sapiens) | HEK293 | ATCC | CRL-1573 | |
Antibody | Anti-HA primary antibody (Rabit monoclonal) | Cell Signaling Technology | Cat.No: 3724. | FCM (1:800) |
Antibody | Goat anti-rabbit IgG(H+L) FITC conjugate secondary antibody (Goat monoclonal) | TransGen Biotech | Cat.No: HS111 | FCM (1:200) |
Recombinant DNA reagent | pCDNA3-HA(plasmid) | This paper | HA version of pCDNA3 | |
Sequence-based reagent | P2Y14-K77A-F | This paper | PCR primers | GACTTTTCCT TTCGCGATCC TTGGTGAC |
Sequence-based reagent | P2Y14-K77A-R | This paper | PCR primers | GTCACCAAGG ATCGCGAAAG GAAAAGTC |
Sequence-based reagent | P2Y14-D81A-F | This paper | PCR primers | CAAGATCCT TGGTGCCTC AGGCCTTGG |
Sequence-based reagent | P2Y14-D81A-R | This paper | PCR primers | GACCAAGGCC TGAGGCACCAA GGATCTTG |
Sequence-based reagent | P2Y14-N90A-F | This paper | PCR primers | GTCCCTGGCA GCTGGCCGTG TTTGTGTGCAG |
Sequence-based reagent | P2Y14-N90A-R | This paper | PCR primers | CTGCACACAAA CACGGCCAGCT GCCAGGGAC |
Sequence-based reagent | P2Y14-I170A-F | This paper | PCR primers | GAGGTTACACA AGCAAAATGTA TAGAACTG |
Sequence-based reagent | P2Y14-I170A-R | This paper | PCR primers | GTTCTATACATT TTGCTTGTGTAA CCTC |
Sequence-based reagent | P2Y14-R274A-F | This paper | PCR primers | CAAAAGAAA TCTTGGCGT ATATGAAAG AATTC |
Sequence-based reagent | P2Y14-R274A-R | This paper | PCR primers | GAATTCTTTC ATATACGCCA AGATTTCTTTT G |
Sequence-based reagent | P2Y14-K277A-F | This paper | PCR primers | CTTGCGGTAT ATGGCAGAAT TCACTCTG |
Sequence-based reagent | P2Y14-K277A-R | This paper | PCR primers | CAGAGTGAAT TCTGCCATAT ACCGCAAG |
Sequence-based reagent | P2Y14-E278A-F | This paper | PCR primers | GCGGTATAT GAAAGCATT CACTCTGCT AC |
Sequence-based reagent | P2Y14-E278A-R | This paper | PCR primers | GTAGCAGAG TGAATGCTTT CATATACCG |
Sequence-based reagent | P2Y12-K80A-F | This paper | PCR primers | CACATTCCC ATTCGCGAT CCTGTCAGA TG |
Sequence-based reagent | P2Y12-K80A-R | This paper | PCR primers | CATCTGACAG GATCGCGAAT GGGAATGTG |
Sequence-based reagent | P2Y12-D84A-F | This paper | PCR primers | CAAGATCCTGT CAGCTGCCAAG CTCGGTAC |
Sequence-based reagent | P2Y12-D84A-R | This paper | PCR primers | GTACCGAGCTT GGCAGCTGACA GGATCTTG |
Sequence-based reagent | P2Y12-F277A-F | This paper | PCR primers | GAGAACACTC TGGCCTACGT CAAGGAATC |
Sequence-based reagent | P2Y12-F277A-R | This paper | PCR primers | GATTCCTTGAC GTAGGCCAGAG TGTTCTC |
Sequence-based reagent | P2Y12-K280A-F | This paper | PCR primers | CTGTTCTACGT CGCGGAATCCA CATTG |
Sequence-based reagent | P2Y12-K280A-R | This paper | PCR primers | CAATGTGGATT CCGCGACGTAG AACAG |
Sequence-based reagent | P2Y12-E281A-F | This paper | PCR primers | GTTCTACGTCA AGGCATCCACA TTGTGGC |
Sequence-based reagent | P2Y12-E281A-R | This paper | PCR primers | GCCACAATGTG GATGCCTTGAC GTAGAAC |
Commercial assay or kit | KOD-plus-Ver.2 | TOYOBO | Cat.No: KOD-211 | |
Commercial assay or kit | MycoBlue Mycoplasma Detector | Vazyme | Cat.No: D101-01 | |
Software, algorithm | GraphPad Prism 6 | GraphPad Prism 6 |
Expression of mutants in HEK293.
Computational characterization of conformational changes and pairwise interactions of simulation models.
Species repertoire information for receptors.