+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-28266 | |||||||||
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Title | Phospholipase C beta 3 (PLCb3) in solution | |||||||||
Map data | final sharpened map | |||||||||
Sample |
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Keywords | PIP2 degradation / IP3 production / DAG production / G protein signaling / HYDROLASE | |||||||||
Function / homology | Function and homology information phosphoinositide phospholipase C / Fatty Acids bound to GPR40 (FFAR1) regulate insulin secretion / Acetylcholine regulates insulin secretion / phosphatidylinositol metabolic process / PLC beta mediated events / regulation of systemic arterial blood pressure / phospholipase C activity / phosphatidylinositol phospholipase C activity / postsynaptic cytosol / Synthesis of IP3 and IP4 in the cytosol ...phosphoinositide phospholipase C / Fatty Acids bound to GPR40 (FFAR1) regulate insulin secretion / Acetylcholine regulates insulin secretion / phosphatidylinositol metabolic process / PLC beta mediated events / regulation of systemic arterial blood pressure / phospholipase C activity / phosphatidylinositol phospholipase C activity / postsynaptic cytosol / Synthesis of IP3 and IP4 in the cytosol / phosphatidylinositol-mediated signaling / lipid catabolic process / release of sequestered calcium ion into cytosol / molecular function activator activity / G beta:gamma signalling through PLC beta / Presynaptic function of Kainate receptors / Ca2+ pathway / phospholipase C-activating G protein-coupled receptor signaling pathway / G alpha (q) signalling events / molecular adaptor activity / calmodulin binding / cadherin binding / G protein-coupled receptor signaling pathway / calcium ion binding / protein-containing complex / membrane / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
Authors | Falzone ME / MacKinnon R | |||||||||
Funding support | United States, 2 items
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Citation | Journal: Proc Natl Acad Sci U S A / Year: 2023 Title: activates hydrolysis by recruiting and orienting on the membrane surface. Authors: Maria E Falzone / Roderick MacKinnon / Abstract: catalyze the hydrolysis of phosphatidylinositol 4, 5-bisphosphate [Formula: see text] into [Formula: see text] [Formula: see text] and [Formula: see text] [Formula: see text]. [Formula: see text] ... catalyze the hydrolysis of phosphatidylinositol 4, 5-bisphosphate [Formula: see text] into [Formula: see text] [Formula: see text] and [Formula: see text] [Formula: see text]. [Formula: see text] regulates the activity of many membrane proteins, while and lead to increased intracellular Ca levels and activate protein kinase C, respectively. are regulated by G protein-coupled receptors through direct interaction with [Formula: see text] and [Formula: see text] and are aqueous-soluble enzymes that must bind to the cell membrane to act on their lipid substrate. This study addresses the mechanism by which [Formula: see text] activates 3. We show that 3 functions as a slow Michaelis-Menten enzyme ( [Formula: see text] ) on membrane surfaces. We used membrane partitioning experiments to study the solution-membrane localization equilibrium of 3. Its partition coefficient is such that only a small quantity of 3 exists in the membrane in the absence of [Formula: see text] . When [Formula: see text] is present, equilibrium binding on the membrane surface increases 3 in the membrane, increasing [Formula: see text] in proportion. Atomic structures on membrane vesicle surfaces show that two [Formula: see text] anchor 3 with its catalytic site oriented toward the membrane surface. Taken together, the enzyme kinetic, membrane partitioning, and structural data show that [Formula: see text] activates by increasing its concentration on the membrane surface and orienting its catalytic core to engage [Formula: see text] . This principle of activation explains rapid stimulated catalysis with low background activity, which is essential to the biological processes mediated by [Formula: see text], and . | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_28266.map.gz | 59.9 MB | EMDB map data format | |
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Header (meta data) | emd-28266-v30.xml emd-28266.xml | 20 KB 20 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_28266_fsc.xml | 9.1 KB | Display | FSC data file |
Images | emd_28266.png | 88.3 KB | ||
Filedesc metadata | emd-28266.cif.gz | 6.6 KB | ||
Others | emd_28266_additional_1.map.gz emd_28266_half_map_1.map.gz emd_28266_half_map_2.map.gz | 49.8 MB 49.8 MB 49.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-28266 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-28266 | HTTPS FTP |
-Validation report
Summary document | emd_28266_validation.pdf.gz | 935.6 KB | Display | EMDB validaton report |
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Full document | emd_28266_full_validation.pdf.gz | 935.2 KB | Display | |
Data in XML | emd_28266_validation.xml.gz | 15.9 KB | Display | |
Data in CIF | emd_28266_validation.cif.gz | 20.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-28266 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-28266 | HTTPS FTP |
-Related structure data
Related structure data | 8emvMC 8emwC 8emxC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_28266.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | final sharpened map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.08 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: final unsharpened map
File | emd_28266_additional_1.map | ||||||||||||
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Annotation | final unsharpened map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_28266_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_28266_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : PLCb3 in solution
Entire | Name: PLCb3 in solution |
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Components |
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-Supramolecule #1: PLCb3 in solution
Supramolecule | Name: PLCb3 in solution / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 138 KDa |
-Macromolecule #1: 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase beta-3
Macromolecule | Name: 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase beta-3 type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: phosphoinositide phospholipase C |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 138.830422 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: GPSRATMALQ LEPPTVVETL RRGSKFIKWD EETSSRNLVT LRVDPNGFFL YWTGPNMEVD TLDISSIRDT RTGRYARLPK DPKIREVLG FGGPDARLEE KLMTVVSGPD PVNTVFLNFM AVQDDTAKVW SEELFKLAMN ILAQNASRNT FLRKAYTKLK L QVNQDGRI ...String: GPSRATMALQ LEPPTVVETL RRGSKFIKWD EETSSRNLVT LRVDPNGFFL YWTGPNMEVD TLDISSIRDT RTGRYARLPK DPKIREVLG FGGPDARLEE KLMTVVSGPD PVNTVFLNFM AVQDDTAKVW SEELFKLAMN ILAQNASRNT FLRKAYTKLK L QVNQDGRI PVKNILKMFS ADKKRVETAL ESCGLKFNRS ESIRPDEFSL EIFERFLNKL CLRPDIDKIL LEIGAKGKPY LT LEQLMDF INQKQRDPRL NEVLYPPLRP SQARLLIEKY EPNQQFLERD QMSMEGFSRY LGGEENGILP LEALDLSTDM TQP LSAYFI NSSHNTYLTA GQLAGTSSVE MYRQALLWGC RCVELDVWKG RPPEEEPFIT HGFTMTTEVP LRDVLEAIAE TAFK TSPYP VILSFENHVD SAKQQAKMAE YCRSIFGDAL LIEPLDKYPL APGVPLPSPQ DLMGRILVKN KKRHRPSAGG PDSAG RKRP LEQSNSALSE SSAATEPSSP QLGSPSSDSC PGLSNGEEVG LEKPSLEPQK SLGDEGLNRG PYVLGPADRE DEEEDE EEE EQTDPKKPTT DEGTASSEVN ATEEMSTLVN YIEPVKFKSF EAARKRNKCF EMSSFVETKA MEQLTKSPME FVEYNKQ QL SRIYPKGTRV DSSNYMPQLF WNVGCQLVAL NFQTLDVAMQ LNAGVFEYNG RSGYLLKPEF MRRPDKSFDP FTEVIVDG I VANALRVKVI SGQFLSDRKV GIYVEVDMFG LPVDTRRKYR TRTSQGNSFN PVWDEEPFDF PKVVLPTLAS LRIAAFEEG GKFVGHRILP VSAIRSGYHY VCLRNEANQP LCLPALLIYT EASDYIPDDH QDYAEALINP IKHVSLMDQR ARQLAALIGE SEAQAGQET CQDTQSQQLG SQPSSNPTPS PLDASPRRPP GPTTSPASTS LSSPGQRDDL IASILSEVAP TPLDELRGHK A LVKLRSRQ ERDLRELRKK HQRKAVTLTR RLLDGLAQAQ AEGRCRLRPG ALGGAADVED TKEGEDEAKR YQEFQNRQVQ SL LELREAQ VDAEAQRRLE HLRQALQRLR EVVLDANTTQ FKRLKEMNER EKKELQKILD RKRHNSISEA KMRDKHKKEA ELT EINRRH ITESVNSIRR LEEAQKQRHD RLVAGQQQVL QQLAEEEPKL LAQLAQECQE QRARLPQEIR RSLLGEMPEG LGDG PLVAC ASNGHAPGSS GHLSGADSES QEENTQL UniProtKB: 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase beta-3 |
-Macromolecule #2: CALCIUM ION
Macromolecule | Name: CALCIUM ION / type: ligand / ID: 2 / Number of copies: 1 / Formula: CA |
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Molecular weight | Theoretical: 40.078 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 4.8 mg/mL |
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Buffer | pH: 7.4 |
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 22 sec. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 289 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 3527 / Average exposure time: 2.0 sec. / Average electron dose: 42.87 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Calibrated defocus min: 1.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |