+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-30047 | |||||||||
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タイトル | CryoEM structure of human PAC1 receptor in complex with maxadilan | |||||||||
マップデータ | ||||||||||
試料 |
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機能・相同性 | 機能・相同性情報 negative regulation of response to reactive oxygen species / development of primary female sexual characteristics / vasoactive intestinal polypeptide receptor activity / positive regulation of cAMP-mediated signaling / NGF-independant TRKA activation / G protein-coupled peptide receptor activity / positive regulation of small GTPase mediated signal transduction / neuropeptide binding / positive regulation of inositol phosphate biosynthetic process / positive regulation of calcium ion transport into cytosol ...negative regulation of response to reactive oxygen species / development of primary female sexual characteristics / vasoactive intestinal polypeptide receptor activity / positive regulation of cAMP-mediated signaling / NGF-independant TRKA activation / G protein-coupled peptide receptor activity / positive regulation of small GTPase mediated signal transduction / neuropeptide binding / positive regulation of inositol phosphate biosynthetic process / positive regulation of calcium ion transport into cytosol / PKA activation in glucagon signalling / peptide hormone binding / adenylate cyclase binding / hair follicle placode formation / developmental growth / D1 dopamine receptor binding / bicellular tight junction / multicellular organismal response to stress / intracellular transport / renal water homeostasis / Hedgehog 'off' state / adenylate cyclase-activating adrenergic receptor signaling pathway / activation of adenylate cyclase activity / cellular response to glucagon stimulus / cAMP-mediated signaling / adenylate cyclase activator activity / regulation of insulin secretion / trans-Golgi network membrane / negative regulation of inflammatory response to antigenic stimulus / bone development / G-protein beta/gamma-subunit complex binding / Olfactory Signaling Pathway / caveola / adenylate cyclase-modulating G protein-coupled receptor signaling pathway / Activation of the phototransduction cascade / adenylate cyclase-activating G protein-coupled receptor signaling pathway / G beta:gamma signalling through PLC beta / Presynaptic function of Kainate receptors / Thromboxane signalling through TP receptor / G protein-coupled acetylcholine receptor signaling pathway / G protein activity / G-protein activation / platelet aggregation / Activation of G protein gated Potassium channels / Inhibition of voltage gated Ca2+ channels via Gbeta/gamma subunits / Prostacyclin signalling through prostacyclin receptor / Glucagon signaling in metabolic regulation / G beta:gamma signalling through CDC42 / cognition / G beta:gamma signalling through BTK / ADP signalling through P2Y purinoceptor 12 / small GTPase binding / Sensory perception of sweet, bitter, and umami (glutamate) taste / Synthesis, secretion, and inactivation of Glucagon-like Peptide-1 (GLP-1) / photoreceptor disc membrane / Glucagon-type ligand receptors / Adrenaline,noradrenaline inhibits insulin secretion / Vasopressin regulates renal water homeostasis via Aquaporins / G alpha (z) signalling events / vasodilation / Glucagon-like Peptide-1 (GLP1) regulates insulin secretion / cellular response to catecholamine stimulus / ADORA2B mediated anti-inflammatory cytokines production / sensory perception of taste / ADP signalling through P2Y purinoceptor 1 / G beta:gamma signalling through PI3Kgamma / adenylate cyclase-activating dopamine receptor signaling pathway / Cooperation of PDCL (PhLP1) and TRiC/CCT in G-protein beta folding / GPER1 signaling / cellular response to prostaglandin E stimulus / Inactivation, recovery and regulation of the phototransduction cascade / G-protein beta-subunit binding / heterotrimeric G-protein complex / sensory perception of smell / G alpha (12/13) signalling events / extracellular vesicle / signaling receptor complex adaptor activity / Thrombin signalling through proteinase activated receptors (PARs) / GTPase binding / response to estradiol / positive regulation of cold-induced thermogenesis / retina development in camera-type eye / signaling receptor activity / Ca2+ pathway / phospholipase C-activating G protein-coupled receptor signaling pathway / G alpha (i) signalling events / fibroblast proliferation / spermatogenesis / G alpha (s) signalling events / 加水分解酵素; 酸無水物に作用; GTPに作用・細胞または細胞小器官の運動に関与 / G alpha (q) signalling events / response to ethanol / Ras protein signal transduction / cell population proliferation / Extra-nuclear estrogen signaling / cell differentiation / receptor complex / cell surface receptor signaling pathway / endosome / response to xenobiotic stimulus 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) / Lutzomyia longipalpis (昆虫) / Lama glama (ラマ) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.6 Å | |||||||||
データ登録者 | Song X / Wang J / Zhang D / Wang HW / Ma Y | |||||||||
引用 | ジャーナル: Cell Res / 年: 2020 タイトル: Cryo-EM structures of PAC1 receptor reveal ligand binding mechanism. 著者: Jia Wang / Xianqiang Song / Dandan Zhang / Xiaoqing Chen / Xun Li / Yaping Sun / Cui Li / Yunpeng Song / Yao Ding / Ruobing Ren / Essa Hu Harrington / Liaoyuan A Hu / Wenge Zhong / Cen Xu / ...著者: Jia Wang / Xianqiang Song / Dandan Zhang / Xiaoqing Chen / Xun Li / Yaping Sun / Cui Li / Yunpeng Song / Yao Ding / Ruobing Ren / Essa Hu Harrington / Liaoyuan A Hu / Wenge Zhong / Cen Xu / Xin Huang / Hong-Wei Wang / Yingli Ma / 要旨: The pituitary adenylate cyclase-activating polypeptide type I receptor (PAC1R) belongs to the secretin receptor family and is widely distributed in the central neural system and peripheral organs. ...The pituitary adenylate cyclase-activating polypeptide type I receptor (PAC1R) belongs to the secretin receptor family and is widely distributed in the central neural system and peripheral organs. Abnormal activation of the receptor mediates trigeminovascular activation and sensitization, which is highly related to migraine, making PAC1R a potential therapeutic target. Elucidation of PAC1R activation mechanism would benefit discovery of therapeutic drugs for neuronal disorders. PAC1R activity is governed by pituitary adenylate cyclase-activating polypeptide (PACAP), known as a major vasodilator neuropeptide, and maxadilan, a native peptide from the sand fly, which is also capable of activating the receptor with similar potency. These peptide ligands have divergent sequences yet initiate convergent PAC1R activity. It is of interest to understand the mechanism of PAC1R ligand recognition and receptor activity regulation through structural biology. Here we report two near-atomic resolution cryo-EM structures of PAC1R activated by PACAP38 or maxadilan, providing structural insights into two distinct ligand binding modes. The structures illustrate flexibility of the extracellular domain (ECD) for ligands with distinct conformations, where ECD accommodates ligands in different orientations while extracellular loop 1 (ECL1) protrudes to further anchor the ligand bound in the orthosteric site. By structure-guided molecular modeling and mutagenesis, we tested residues in the ligand-binding pockets and identified clusters of residues that are critical for receptor activity. The structures reported here for the first time elucidate the mechanism of specificity and flexibility of ligand recognition and binding for PAC1R, and provide insights toward the design of therapeutic molecules targeting PAC1R. | |||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_30047.map.gz | 2.3 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-30047-v30.xml emd-30047.xml | 17.1 KB 17.1 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_30047.png | 81 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-30047 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-30047 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_30047_validation.pdf.gz | 329 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_30047_full_validation.pdf.gz | 328.5 KB | 表示 | |
XML形式データ | emd_30047_validation.xml.gz | 5.5 KB | 表示 | |
CIF形式データ | emd_30047_validation.cif.gz | 6.2 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-30047 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-30047 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_30047.map.gz / 形式: CCP4 / 大きさ: 22.2 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.091 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-試料の構成要素
-全体 : PACAP38-PAC1R complex
全体 | 名称: PACAP38-PAC1R complex |
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要素 |
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-超分子 #1: PACAP38-PAC1R complex
超分子 | 名称: PACAP38-PAC1R complex / タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: all |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
組換発現 | 生物種: Spodoptera frugiperda (ツマジロクサヨトウ) |
分子量 | 理論値: 150 KDa |
-分子 #1: Pituitary adenylate cyclase-activating polypeptide type I receptor
分子 | 名称: Pituitary adenylate cyclase-activating polypeptide type I receptor タイプ: protein_or_peptide / ID: 1 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 47.967223 KDa |
組換発現 | 生物種: Insect BA phytoplasma (バクテリア) |
配列 | 文字列: DYKDDDDKIF KKEQAMCLEK IQRANELMGF NDSSPGCPGM WDNITCWKPA HVGEMVLVSC PELFRIFNPD QDMGVVSRNC TEDGWSEPF PHYFDACGFD EYESETGDQD YYYLSVKALY TVGYSLSLVA LLLAMVILCR FRKLHCTRNF IHMNLFVSFM L RAISVFIK ...文字列: DYKDDDDKIF KKEQAMCLEK IQRANELMGF NDSSPGCPGM WDNITCWKPA HVGEMVLVSC PELFRIFNPD QDMGVVSRNC TEDGWSEPF PHYFDACGFD EYESETGDQD YYYLSVKALY TVGYSLSLVA LLLAMVILCR FRKLHCTRNF IHMNLFVSFM L RAISVFIK DWILYAEQDS NHCFISTVEC KAVMVFFHYC VVSNYFWLFI EGLYLFTLLV ETFFPERRYF YWYTIIGWGA PL VFVTVWA TLRLYFDDTG CWDMNDSTAL WWVIKGPVVG SIMVNFVLFI GIIVILVQKL QSPDMGGNES SIYLRLARST LLL IPLFGI HYTVFAFSPE NVSKRERLVF ELGLGSFQGF VVAVLYCFLN GEVQAEIKRK WRSWKVNRYF AVDFKHRHPS LASS HHHHH HHHHH |
-分子 #2: Maxadilan
分子 | 名称: Maxadilan / タイプ: protein_or_peptide / ID: 2 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: Lutzomyia longipalpis (昆虫) |
分子量 | 理論値: 6.88182 KDa |
配列 | 文字列: CDATCQFRKA IDDCQKQAHH SNVLQTSVQT TATFTSMDTS QLPGNSVFKE CMKQKKKEFK A |
-分子 #3: Nanobody 35
分子 | 名称: Nanobody 35 / タイプ: protein_or_peptide / ID: 3 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: Lama glama (ラマ) |
分子量 | 理論値: 14.71432 KDa |
組換発現 | 生物種: Escherichia coli (大腸菌) |
配列 | 文字列: QVQLQESGGG LVQPGGSLRL SCAASGFTFS NYKMNWVRQA PGKGLEWVSD ISQSGASISY TGSVKGRFTI SRDNAKNTLY LQMNSLKPE DTAVYYCARC PAPFTRDCFD VTSTTYAYRG QGTQVTVSSH HHHHH |
-分子 #4: Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2
分子 | 名称: Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2 タイプ: protein_or_peptide / ID: 4 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 7.861143 KDa |
組換発現 | 生物種: Insect BA phytoplasma (バクテリア) |
配列 | 文字列: MASNNTASIA QARKLVEQLK MEANIDRIKV SKAAADLMAY CEAHAKEDPL LTPVPASENP FREKKFFCAI L |
-分子 #5: Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1
分子 | 名称: Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1 タイプ: protein_or_peptide / ID: 5 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 37.47398 KDa |
組換発現 | 生物種: Insect BA phytoplasma (バクテリア) |
配列 | 文字列: GMSELDQLRQ EAEQLKNQIR DARKACADAT LSQITNNIDP VGRIQMRTRR TLRGHLAKIY AMHWGTDSRL LVSASQDGKL IIWDSYTTN KVHAIPLRSS WVMTCAYAPS GNYVACGGLD NICSIYNLKT REGNVRVSRE LAGHTGYLSC CRFLDDNQIV T SSGDTTCA ...文字列: GMSELDQLRQ EAEQLKNQIR DARKACADAT LSQITNNIDP VGRIQMRTRR TLRGHLAKIY AMHWGTDSRL LVSASQDGKL IIWDSYTTN KVHAIPLRSS WVMTCAYAPS GNYVACGGLD NICSIYNLKT REGNVRVSRE LAGHTGYLSC CRFLDDNQIV T SSGDTTCA LWDIETGQQT TTFTGHTGDV MSLSLAPDTR LFVSGACDAS AKLWDVREGM CRQTFTGHES DINAICFFPN GN AFATGSD DATCRLFDLR ADQELMTYSH DNIICGITSV SFSKSGRLLL AGYDDFNCNV WDALKADRAG VLAGHDNRVS CLG VTDDGM AVATGSWDSF LKIWN |
-分子 #6: Guanine nucleotide-binding protein G(s) subunit alpha isoforms short
分子 | 名称: Guanine nucleotide-binding protein G(s) subunit alpha isoforms short タイプ: protein_or_peptide / ID: 6 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 45.683434 KDa |
組換発現 | 生物種: Insect BA phytoplasma (バクテリア) |
配列 | 文字列: MGCLGNSKTE DQRNEEKAQR EANKKIEKQL QKDKQVYRAT HRLLLLGAGE SGKNTIVKQM RILHVNGFNG EGGEEDPQAA RSNSDGEKA TKVQDIKNNL KEAIETIVAA MSNLVPPVEL ANPENQFRVD YILSVMNVPD FDFPPEFYEH AKALWEDEGV R ACYERSNE ...文字列: MGCLGNSKTE DQRNEEKAQR EANKKIEKQL QKDKQVYRAT HRLLLLGAGE SGKNTIVKQM RILHVNGFNG EGGEEDPQAA RSNSDGEKA TKVQDIKNNL KEAIETIVAA MSNLVPPVEL ANPENQFRVD YILSVMNVPD FDFPPEFYEH AKALWEDEGV R ACYERSNE YQLIDCAQYF LDKIDVIKQA DYVPSDQDLL RCRVLTSGIF ETKFQVDKVN FHMFDVGAQR DERRKWIQCF ND VTAIIFV VASSSYNMVI REDNQTNRLQ AALKLFDSIW NNKWLRDTSV ILFLNKQDLL AEKVLAGKSK IEDYFPEFAR YTT PEDATP EPGEDPRVTR AKYFIRDEFL RISTASGDGR HYCYPHFTCA VDTENIRRVF NDCRDIIQRM HLRQYELL |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 5 mg/mL |
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緩衝液 | pH: 7.5 |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: FEI VITROBOT MARK IV |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: SUPER-RESOLUTION / デジタル化 - サイズ - 横: 3838 pixel / デジタル化 - サイズ - 縦: 3710 pixel / 平均電子線量: 50.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 50.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 0.0 mm |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
CTF補正 | ソフトウェア - 名称: CTFFIND (ver. 4.1) |
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最終 再構成 | 使用したクラス数: 1 / 想定した対称性 - 点群: C1 (非対称) / アルゴリズム: BACK PROJECTION / 解像度のタイプ: BY AUTHOR / 解像度: 3.6 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: RELION (ver. 3.0) / 使用した粒子像数: 58451 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |