+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-27344 | |||||||||
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タイトル | cryo-EM structure of TRPM3 ion channel in complex with Gbg, tethered by ALFA-nanobody | |||||||||
マップデータ | sharpened map with a B-factor of 112.4 | |||||||||
試料 |
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キーワード | TRPM3 / ion channel / Gbg / MEMBRANE PROTEIN | |||||||||
機能・相同性 | 機能・相同性情報 metal ion transport / monoatomic cation transmembrane transport / monoatomic cation transport / monoatomic cation channel activity / protein tetramerization / Olfactory Signaling Pathway / Activation of the phototransduction cascade / G beta:gamma signalling through PLC beta / Presynaptic function of Kainate receptors / Thromboxane signalling through TP receptor ...metal ion transport / monoatomic cation transmembrane transport / monoatomic cation transport / monoatomic cation channel activity / protein tetramerization / Olfactory Signaling Pathway / Activation of the phototransduction cascade / 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 activation / 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 / ADP signalling through P2Y purinoceptor 12 / G beta:gamma signalling through BTK / Synthesis, secretion, and inactivation of Glucagon-like Peptide-1 (GLP-1) / Sensory perception of sweet, bitter, and umami (glutamate) taste / photoreceptor disc membrane / Adrenaline,noradrenaline inhibits insulin secretion / Glucagon-type ligand receptors / Vasopressin regulates renal water homeostasis via Aquaporins / G alpha (z) signalling events / cellular response to catecholamine stimulus / Glucagon-like Peptide-1 (GLP1) regulates insulin secretion / ADORA2B mediated anti-inflammatory cytokines production / sensory perception of taste / ADP signalling through P2Y purinoceptor 1 / adenylate cyclase-activating dopamine receptor signaling pathway / G beta:gamma signalling through PI3Kgamma / cellular response to prostaglandin E stimulus / Cooperation of PDCL (PhLP1) and TRiC/CCT in G-protein beta folding / GPER1 signaling / G-protein beta-subunit binding / Inactivation, recovery and regulation of the phototransduction cascade / heterotrimeric G-protein complex / G alpha (12/13) signalling events / extracellular vesicle / signaling receptor complex adaptor activity / Thrombin signalling through proteinase activated receptors (PARs) / GTPase binding / retina development in camera-type eye / Ca2+ pathway / phospholipase C-activating G protein-coupled receptor signaling pathway / G alpha (i) signalling events / fibroblast proliferation / G alpha (s) signalling events / G alpha (q) signalling events / Ras protein signal transduction / Extra-nuclear estrogen signaling / cell population proliferation / G protein-coupled receptor signaling pathway / lysosomal membrane / GTPase activity / synapse / protein-containing complex binding / signal transduction / extracellular exosome / membrane / plasma membrane / cytoplasm / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | Mus musculus (ハツカネズミ) / Homo sapiens (ヒト) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.7 Å | |||||||||
データ登録者 | Zhao C / MacKinnon R | |||||||||
資金援助 | 米国, 1件
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引用 | ジャーナル: Neuron / 年: 2023 タイトル: Structural and functional analyses of a GPCR-inhibited ion channel TRPM3. 著者: Chen Zhao / Roderick MacKinnon / 要旨: G-protein coupled receptors (GPCRs) govern the physiological response to stimuli by modulating the activity of downstream effectors, including ion channels. TRPM3 is an ion channel inhibited by GPCRs ...G-protein coupled receptors (GPCRs) govern the physiological response to stimuli by modulating the activity of downstream effectors, including ion channels. TRPM3 is an ion channel inhibited by GPCRs through direct interaction with G protein (Gβγ) released upon their activation. This GPCR-TRPM3 signaling pathway contributes to the analgesic effect of morphine. Here, we characterized Gβγ inhibition of TRPM3 using electrophysiology and single particle cryo-electron microscopy (cryo-EM). From electrophysiology, we obtained a half inhibition constant (IC50) of ∼240 nM. Using cryo-EM, we determined structures of mouse TRPM3 expressed in human cells with and without Gβγ and with and without PIP, a lipid required for TRPM3 activity, at resolutions of 2.7-4.7 Å. Gβγ-TRPM3 interfaces vary depending on PIP occupancy; however, in all cases, Gβγ appears loosely attached to TRPM3. The IC50 in electrophysiology experiments raises the possibility that additional unknown factors may stabilize the TRPM3-Gβγ complex. | |||||||||
履歴 |
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-構造の表示
添付画像 |
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-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_27344.map.gz | 16.2 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-27344-v30.xml emd-27344.xml | 17.8 KB 17.8 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_27344_fsc.xml | 15.9 KB | 表示 | FSCデータファイル |
画像 | emd_27344.png | 101.6 KB | ||
Filedesc metadata | emd-27344.cif.gz | 6.6 KB | ||
その他 | emd_27344_half_map_1.map.gz emd_27344_half_map_2.map.gz | 275.5 MB 275.5 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-27344 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-27344 | HTTPS FTP |
-関連構造データ
関連構造データ | 8ddwMC 8ddqC 8ddrC 8ddsC 8ddtC 8dduC 8ddvC 8ddxC 8ed7C 8ed8C 8ed9C M: このマップから作成された原子モデル C: 同じ文献を引用 (文献) |
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類似構造データ | 類似検索 - 機能・相同性F&H 検索 |
-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_27344.map.gz / 形式: CCP4 / 大きさ: 343 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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注釈 | sharpened map with a B-factor of 112.4 | ||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.08 Å | ||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-ハーフマップ: half map 1
ファイル | emd_27344_half_map_1.map | ||||||||||||
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注釈 | half map 1 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: half map 2
ファイル | emd_27344_half_map_2.map | ||||||||||||
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注釈 | half map 2 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
-全体 : TRPM3
全体 | 名称: TRPM3 |
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要素 |
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-超分子 #1: TRPM3
超分子 | 名称: TRPM3 / 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: Mus musculus (ハツカネズミ) |
-分子 #1: Transient receptor potential cation channel, subfamily M, member 3
分子 | 名称: Transient receptor potential cation channel, subfamily M, member 3 タイプ: protein_or_peptide / ID: 1 / コピー数: 4 / 光学異性体: LEVO |
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由来(天然) | 生物種: Mus musculus (ハツカネズミ) |
分子量 | 理論値: 157.812922 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: MGKKWRDAGE LERGCSDRED SAESRRRSRS ASRGRFAESW KRLSSKQGST KRSGLPAQQT PAQKSWIERA FYKRECVHII PSTKDPHRC CCGRLIGQHV GLTPSISVLQ NEKNESRLSR NDIQSEKWSI SKHTQLSPTD AFGTIEFQGG GHSNKAMYVR V SFDTKPDL ...文字列: MGKKWRDAGE LERGCSDRED SAESRRRSRS ASRGRFAESW KRLSSKQGST KRSGLPAQQT PAQKSWIERA FYKRECVHII PSTKDPHRC CCGRLIGQHV GLTPSISVLQ NEKNESRLSR NDIQSEKWSI SKHTQLSPTD AFGTIEFQGG GHSNKAMYVR V SFDTKPDL LLHLMTKEWQ LELPKLLISV HGGLQNFELQ PKLKQVFGKG LIKAAMTTGA WIFTGGVNTG VIRHVGDALK DH ASKSRGK ICTIGIAPWG IVENQEDLIG RDVVRPYQTM SNPMSKLTVL NSMHSHFILA DNGTTGKYGA EVKLRRQLEK HIS LQKINT RIGQGVPVVA LIVEGGPNVI SIVLEYLRDT PPVPVVVCDG SGRASDILAF GHKYSEEGGL INESLRDQLL VTIQ KTFTY TRTQAQHLFI ILMECMKKKE LITVFRMGSE GHQDIDLAIL TALLKGANAS APDQLSLALA WNRVDIARSQ IFIYG QQWP VGSLEQAMLD ALVLDRVDFV KLLIENGVSM HRFLTISRLE ELYNTRHGPS NTLYHLVRDV KKGNLPPDYR ISLIDI GLV IEYLMGGAYR CNYTRKRFRT LYHNLFGPKR PKALKLLGME DDIPLRRGRK TTKKREEEVD IDLDDPEINH FPFPFHE LM VWAVLMKRQK MALFFWQHGE EAMAKALVAC KLCKAMAHEA SENDMVDDIS QELNHNSRDF GQLAVELLDQ SYKQDEQL A MKLLTYELKN WSNATCLQLA VAAKHRDFIA HTCSQMLLTD MWMGRLRMRK NSGLKVILGI LLPPSILSLE FKNKDDMPY MTQAQEIHLQ EKEPEEPEKP TKEKDEEDME LTAMLGRSNG ESSRKKDEEE VQSRHRLIPV GRKIYEFYNA PIVKFWFYTL AYIGYLMLF NYIVLVKMER WPSTQEWIVI SYIFTLGIEK MREILMSEPG KLLQKVKVWL QEYWNVTDLI AILLFSVGMI L RLQDQPFR SDGRVIYCVN IIYWYIRLLD IFGVNKYLGP YVMMIGKMMI DMMYFVIIML VVLMSFGVAR QAILFPNEEP SW KLAKNIF YMPYWMIYGE VFADQIDPPC GQNETREDGK TIQLPPCKTG AWIVPAIMAC YLLVANILLV NLLIAVFNNT FFE VKSISN QVWKFQRYQL IMTFHERPVL PPPLIIFSHM TMIFQHVCCR WRKHESDQDE RDYGLKLFIT DDELKKVHDF EEQC IEEYF REKDDRFNSS NDERIRVTSE RVENMSMRLE EVNEREHSMK ASLQTVDIRL AQLEDLIGRM ATALERLTGL ERAES NKIR SRTSSDCTDA AYIVRQSSFN SQEGNTFKLQ ESIDPAGEET ISPTSPTLMP RMRSHSFYSV NVKDKGGIEK LESIFK ERS LSLHRATS UniProtKB: Transient receptor potential cation channel, subfamily M, member 3 |
-分子 #2: Unidentified segment at the N-terminus of TRPM3
分子 | 名称: Unidentified segment at the N-terminus of TRPM3 / タイプ: protein_or_peptide / ID: 2 詳細: a segment at the N-terminus of TRPM3 whose sequence cannot be identified from the cryo-EM density コピー数: 4 / 光学異性体: LEVO |
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由来(天然) | 生物種: Mus musculus (ハツカネズミ) |
分子量 | 理論値: 1.464797 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) |
-分子 #3: 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: 3 / 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: 37.285734 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: SELDQLRQEA EQLKNQIRDA RKACADATLS QITNNIDPVG RIQMRTRRTL RGHLAKIYAM HWGTDSRLLV SASQDGKLII WDSYTTNKV HAIPLRSSWV MTCAYAPSGN YVACGGLDNI CSIYNLKTRE GNVRVSRELA GHTGYLSCCR FLDDNQIVTS S GDTTCALW ...文字列: SELDQLRQEA EQLKNQIRDA RKACADATLS QITNNIDPVG RIQMRTRRTL RGHLAKIYAM HWGTDSRLLV SASQDGKLII WDSYTTNKV HAIPLRSSWV MTCAYAPSGN YVACGGLDNI CSIYNLKTRE GNVRVSRELA GHTGYLSCCR FLDDNQIVTS S GDTTCALW DIETGQQTTT FTGHTGDVMS LSLAPDTRLF VSGACDASAK LWDVREGMCR QTFTGHESDI NAICFFPNGN AF ATGSDDA TCRLFDLRAD QELMTYSHDN IICGITSVSF SKSGRLLLAG YDDFNCNVWD ALKADRAGVL AGHDNRVSCL GVT DDGMAV ATGSWDSFLK IWN UniProtKB: Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1 |
-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: 7.713882 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: ASNNTASIAQ ARKLVEQLKM EANIDRIKVS KAAADLMAYC EAHAKEDPLL TPVPASENPF REKKFFSAIL UniProtKB: Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2 |
-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
緩衝液 | pH: 8.5 |
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凍結 | 凍結剤: ETHANE |
-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 K3 (6k x 4k) / 平均電子線量: 51.4 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.0 µm / 最小 デフォーカス(公称値): 1.0 µm |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |