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2021-06-18T13:36:36+09:00
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Ritsumeikan Univ.KO-325
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(Discovery of surrogate ligands for a GPCR based on the 3D receptor model\
.)Tj
/TT1 1 Tf
12 0 0 12 475.0234 759.75 Tm
(Takeshi Hiramoto)Tj
-35.877 -2.313 Td
(G-protein coupled receptors \(GPCRs\) play crucial roles in many physiol\
ogical functions in the whole)Tj
-1 -1.5 Td
(human body, so that GPCRs represent one of the most critical classes of \
confirmed drug targets. The)Tj
0 -1.5 TD
(elucidation of the 3D structures of GPCRs is indispensable for novel dru\
g design. However, since a)Tj
T*
(membrane bound protein such as a GPCR is not easily crystallized, an X-r\
ay crystallography analysis cannot)Tj
T*
(be expected. Therefore, I have conducted )Tj
/TT2 1 Tf
16.769 0 Td
(in silico)Tj
/TT1 1 Tf
3.194 0 Td
( and )Tj
/TT2 1 Tf
(in vitro)Tj
/TT1 1 Tf
4.86 0 Td
( studies to elucidate a 3D structure of a ligand-)Tj
-24.823 -1.5 Td
(docked GPCR and to develop a new biorational approach for the drug disco\
very using the 3D structure.)Tj
1 -2.5 Td
(First, I constructed the 3D structure of bovine rhodopsin by using Fouri\
er transform analysis and a)Tj
-1 -1.5 Td
(homology modeling. I found that the 3D model showed a good agreement wit\
h the reported retinal binding)Tj
0 -1.562 TD
(\(Section I\). Using the similar method, I constructed the model structu\
re of the human P2Y)Tj
8.25 0 0 8.25 465.7363 576.75 Tm
(1)Tj
12 0 0 12 469.8613 575.25 Tm
( receptor, one of)Tj
-36.447 -1.5 Td
(the GPCRs. Then, I made the docking model of ADP and hP2Y1 receptor comp\
lex. The model showed that)Tj
0 -1.5 TD
(ADP reasonably bound inside the helical bundle known to an agonist and a\
ntagonist binding sites. Then I)Tj
0 -1.562 TD
(employed an )Tj
/TT2 1 Tf
5.387 0 Td
(in silico)Tj
/TT1 1 Tf
3.194 0 Td
( screening for endogenous compounds to select possible hP2Y)Tj
8.25 0 0 8.25 435.4258 522 Tm
(1)Tj
12 0 0 12 439.5508 520.5 Tm
( ligands, by using)Tj
-33.921 -1.5 Td
(AutoDock 3.0. As a result, 21 compounds were selected including known P2\
Y1 agonists and antagonists)Tj
0 -1.5 TD
(\(Section II\).)Tj
1 -2.5 Td
(I have prepared recombinant CHO cells expressing hP2Y1 \(or hP2Y2\) rece\
ptors. The cells were clearly)Tj
-1 -1.562 Td
(shown to increase in intracellular Ca)Tj
8.25 0 0 8.25 208.1055 437.25 Tm
(2+)Tj
12 0 0 12 216.8832 435.75 Tm
( concentration \([Ca)Tj
8.25 0 0 8.25 309.5082 437.25 Tm
(2+)Tj
12 0 0 12 318.2859 435.75 Tm
(]i\) by ADP at a concentration of 10)Tj
8.25 0 0 8.25 487.9031 437.25 Tm
(-6)Tj
/C0_0 1 Tf
0.833 -0.091 Td
<0299>Tj
/TT1 1 Tf
1 0.091 Td
(-9)Tj
12 0 0 12 509.8977 435.75 Tm
(M. The)Tj
-39.783 -1.562 Td
(ADP-induced Ca)Tj
8.25 0 0 8.25 114.8184 418.5 Tm
(2+)Tj
12 0 0 12 123.5961 417 Tm
( response was blocked by the tentative P2Y1 antagonist, 3'P5'P adenosine\
\(Section III\).)Tj
-7.591 -1.5 Td
(Using this assay, I have evaluated the 21 compounds mentioned above, and\
identified 3 noble compounds that)Tj
0 -1.562 TD
(increased the [Ca)Tj
8.25 0 0 8.25 115.791 381.75 Tm
(2+)Tj
12 0 0 12 124.5687 380.25 Tm
(]i through activating the hP2Y)Tj
8.25 0 0 8.25 270.5433 381.75 Tm
(1)Tj
12 0 0 12 274.6683 380.25 Tm
( receptor stably expressed in recombinant CHO cells.)Tj
-20.181 -1.562 Td
(Among them, 5-phosphoribosyl 1-pyrophosphate \(PRPP\) activated the hP2Y\
)Tj
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(1)Tj
12 0 0 12 406.5918 361.5 Tm
( receptor with the lowest ED)Tj
8.25 0 0 8.25 544.2168 363 Tm
(50)Tj
12 0 0 12 32.5 343.5 Tm
(value of 15nM. PRPP was shown to specifically activate hP2Y1 receptor.)Tj
1 -2.563 Td
(This is the first report that demonstrates the surrogate ligands for the\
hP2Y)Tj
8.25 0 0 8.25 403.7559 314.25 Tm
(1)Tj
12 0 0 12 407.8809 312.75 Tm
( receptor were identified by)Tj
-31.282 -1.5 Td
(using )Tj
/TT2 1 Tf
(in silico)Tj
/TT1 1 Tf
5.611 0 Td
( and )Tj
/TT2 1 Tf
(in vitro)Tj
/TT1 1 Tf
4.86 0 Td
( studies. Accumulation of these data can make it possible to refine the \
3D structure)Tj
-10.472 -1.5 Td
(of P2Y receptor as well as to develop a computational method to elucidat\
e the mechanism by which a ligand)Tj
0 -1.5 TD
(binds to a GPCR. In addition, the usage of these approaches described he\
re should be expected to discover)Tj
T*
(orphan GPCR ligands.)Tj
ET
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