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- EMDB-16252: Structure of Adenylyl cyclase 8 bound to stimulatory G-protein, C... -
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Basic information
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Title | Structure of Adenylyl cyclase 8 bound to stimulatory G-protein, Ca2+/Calmodulin, Forskolin and MANT-GTP | |||||||||
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![]() | Adenylyl Cyclase / cAMP signaling / G proteins / Calmodulin / MEMBRANE PROTEIN | |||||||||
Function / homology | ![]() cellular response to morphine / Adenylate cyclase activating pathway / Adenylate cyclase inhibitory pathway / positive regulation of long-term synaptic depression / glucose mediated signaling pathway / calcium- and calmodulin-responsive adenylate cyclase activity / regulation of cellular response to stress / sensory perception of chemical stimulus / PKA activation / neuroinflammatory response ...cellular response to morphine / Adenylate cyclase activating pathway / Adenylate cyclase inhibitory pathway / positive regulation of long-term synaptic depression / glucose mediated signaling pathway / calcium- and calmodulin-responsive adenylate cyclase activity / regulation of cellular response to stress / sensory perception of chemical stimulus / PKA activation / neuroinflammatory response / adenylate cyclase / positive regulation of synaptic plasticity / mu-type opioid receptor binding / Hedgehog 'off' state / corticotropin-releasing hormone receptor 1 binding / cAMP biosynthetic process / adenylate cyclase activity / G alpha (z) signalling events / beta-2 adrenergic receptor binding / neuronal cell body membrane / cellular response to glucagon stimulus / presynaptic active zone / positive regulation of insulin secretion involved in cellular response to glucose stimulus / excitatory synapse / D1 dopamine receptor binding / long-term memory / adenylate cyclase-activating adrenergic receptor signaling pathway / regulation of cytosolic calcium ion concentration / insulin-like growth factor receptor binding / adenylate cyclase activator activity / ionotropic glutamate receptor binding / hippocampal mossy fiber to CA3 synapse / positive regulation of long-term synaptic potentiation / locomotory behavior / G-protein beta/gamma-subunit complex binding / Schaffer collateral - CA1 synapse / adenylate cyclase-activating G protein-coupled receptor signaling pathway / adenylate cyclase-activating dopamine receptor signaling pathway / heterotrimeric G-protein complex / presynaptic membrane / basolateral plasma membrane / postsynaptic density / intracellular signal transduction / apical plasma membrane / axon / GTPase activity / glutamatergic synapse / dendrite / GTP binding / ATP binding / metal ion binding / plasma membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.5 Å | |||||||||
![]() | Khanppnavar B / Korkhov VM / Mehta V | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Regulatory sites of CaM-sensitive adenylyl cyclase AC8 revealed by cryo-EM and structural proteomics. Authors: Basavraj Khanppnavar / Dina Schuster / Pia Lavriha / Federico Uliana / Merve Özel / Ved Mehta / Alexander Leitner / Paola Picotti / Volodymyr M Korkhov / ![]() Abstract: Membrane adenylyl cyclase AC8 is regulated by G proteins and calmodulin (CaM), mediating the crosstalk between the cAMP pathway and Ca signalling. Despite the importance of AC8 in physiology, the ...Membrane adenylyl cyclase AC8 is regulated by G proteins and calmodulin (CaM), mediating the crosstalk between the cAMP pathway and Ca signalling. Despite the importance of AC8 in physiology, the structural basis of its regulation by G proteins and CaM is not well defined. Here, we report the 3.5 Å resolution cryo-EM structure of the bovine AC8 bound to the stimulatory Gαs protein in the presence of Ca/CaM. The structure reveals the architecture of the ordered AC8 domains bound to Gαs and the small molecule activator forskolin. The extracellular surface of AC8 features a negatively charged pocket, a potential site for unknown interactors. Despite the well-resolved forskolin density, the captured state of AC8 does not favour tight nucleotide binding. The structural proteomics approaches, limited proteolysis and crosslinking mass spectrometry (LiP-MS and XL-MS), allowed us to identify the contact sites between AC8 and its regulators, CaM, Gαs, and Gβγ, as well as to infer the conformational changes induced by these interactions. Our results provide a framework for understanding the role of flexible regions in the mechanism of AC regulation. | |||||||||
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 18.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 18.8 KB 18.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 15 KB | Display | ![]() |
Images | ![]() | 123.2 KB | ||
Filedesc metadata | ![]() | 6.7 KB | ||
Others | ![]() ![]() | 259.9 MB 255.1 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8buzMC ![]() 8bv5C M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Voxel size | X=Y=Z: 0.66 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: half2
File | emd_16252_half_map_1.map | ||||||||||||
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Annotation | half2 | ||||||||||||
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-Half map: half1
File | emd_16252_half_map_2.map | ||||||||||||
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Annotation | half1 | ||||||||||||
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Sample components
-Entire : Structure of Adenylyl cyclase 8 bound to stimulatory G protein, C...
Entire | Name: Structure of Adenylyl cyclase 8 bound to stimulatory G protein, Ca2+/Calmodulin, Forskolin, and MANT-GTP |
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Components |
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-Supramolecule #1: Structure of Adenylyl cyclase 8 bound to stimulatory G protein, C...
Supramolecule | Name: Structure of Adenylyl cyclase 8 bound to stimulatory G protein, Ca2+/Calmodulin, Forskolin, and MANT-GTP type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 |
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-Supramolecule #2: Adenylyl cyclase 8
Supramolecule | Name: Adenylyl cyclase 8 / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() ![]() |
-Supramolecule #3: G protein alpha S
Supramolecule | Name: G protein alpha S / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2 |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #1: Adenylate cyclase type 8
Macromolecule | Name: Adenylate cyclase type 8 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: adenylate cyclase |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 140.192328 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MELSDVRCLS GSEELYTIHP TPPAGDSGSG SRPQRLLWQT AVRHITEQRF IHGHRGGGGG GGNGSSSKAS DPGGGGPNHH HASQLSGDS ALPLYALGPG ERAHGTGGPK VFPERSGSGS ASGSGGGGDL GFLHLDCAPS NSDFFLNGGY SYRGVIFPTL R NSFKSRDL ...String: MELSDVRCLS GSEELYTIHP TPPAGDSGSG SRPQRLLWQT AVRHITEQRF IHGHRGGGGG GGNGSSSKAS DPGGGGPNHH HASQLSGDS ALPLYALGPG ERAHGTGGPK VFPERSGSGS ASGSGGGGDL GFLHLDCAPS NSDFFLNGGY SYRGVIFPTL R NSFKSRDL ERLYQRYFLG QRRKSEVVMN VLDVLTKLTL LVLHLSLASA PMDPLKGILL GFFTGIEVVI CALVVVRKDT TS HTYLQYS GVVTWVAMTT QILAAGLGYG LLGDGIGYVL FTLFATYSML PLPLTWAILA GLGTSLMQVV LQAVIPRLAV ISI NQVVAQ AVLFMCMNTA GIFISYLSDR AQRQAFLETR RCVEARLRLE TENQRQERLV LSVLPRFVVL EMINDMTNVE DEHL QHQFH RIYIHRYENV SILFADVKGF TNLSTTLSAQ ELVRMLNELF ARFDRLAHEH HCLRIKILGD CYYCVSGLPE PRQDH AHCC VEMGLSMIKT IRYVRSRTKH DVDMRIGIHS GSVLCGVLGL RKWQFDVWSW DVDIANKLES GGIPGRIHIS KATLDC LNG DYNVEEGHGK ERNEFLRKHN IETYLIKQPE ESLLSLPEDI VKESVSSSDR RNSGATFTEG SWSPELPFDN IVGKQNT LA ALTRNSINLL PNHLAQALHV QSGPEEINKR IEHTIDLRSG DKLRREHIKP FSLMFKDSSL EHKYSQMRDE VFKSNLVC A FIVLLFITAI QSLLPSSRVM PMAIQFSILI MLHSALVLIT TAEDYKCLPL VLRKTCCWIN ETYLARNVII FASILINFL GAILNILWCD FDKSIPLKNL TFNSSAVFTD ICSYPEYFVF TGVLAMVTCA VFLRLNSVLK LAVLLIMIAI YALLTETIYA GLFLRYDNL NHSGEDFLGT KEASLLLMAM FLLAVFYHGQ QLEYTARLDF LWRVQAKEEI NEMKELREHN ENMLRNILPS H VARHFLEK DRDNEELYSQ SYDAVGVMFA SIPGFADFYS QTEMNNQGVE CLRLLNEIIA DFDELLGEDR FQDIEKIKTI GS TYMAVSG LSPEKQQCED KWGHLCALAD FSLALTESIQ EINKHSFNNF ELRIGISHGS VVAGVIGAKK PQYDIWGKTV NLA SRMDST GVSGRIQVPE ETYLILKDQG FAFDYRGEIY VKGISEQEGK IKTYFLLGRV QPNPFILPPR RLPGQYSLAA VVLG LVQSL NRQRQKQLLN ENNNTGIIKG HYNRRTLLTP SGPEPGAPAE GADKPELP UniProtKB: Adenylate cyclase type 8 |
-Macromolecule #2: Guanine nucleotide-binding protein G(s) subunit alpha isoforms short
Macromolecule | Name: Guanine nucleotide-binding protein G(s) subunit alpha isoforms short type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 47.003801 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MGCLGNSKTE DQRNEEKAQR EANKKIEKQL QKDKQVYRAT HRLLLLGAGE SGKSTIVKQM RILHVNGFNG GEGGEEDPNA AKSNSDGEK ATKVQDIKNN LKEAIETIVA AMSNLVPPVE LANPENQFRV DYILSVMNVP DFDFPPEFYE HAKALWEDEG V RACYERSN ...String: MGCLGNSKTE DQRNEEKAQR EANKKIEKQL QKDKQVYRAT HRLLLLGAGE SGKSTIVKQM RILHVNGFNG GEGGEEDPNA AKSNSDGEK ATKVQDIKNN LKEAIETIVA AMSNLVPPVE LANPENQFRV DYILSVMNVP DFDFPPEFYE HAKALWEDEG V RACYERSN EYQLIDCAQY FLDKIDVIKQ DDYVPSDQDL LRCRVLTSGI FETKFQVDKV NFHMFDVGGQ RDERRKWIQC FN DVTAIIF VVASSSYNMV IREDNQTNRL QEALNLFKSI WNNRWLRTIS VILFLNKQDL LAEKVLAGKS KIEDYFPEFA RYT TPEDAT PEPGEDPRVT RAKYFIRDEF LRISTASGDG RHYCYPHFTC AVDTENIRRV FNDCRDIIQR MHLRQYELLG GHHH HHHHH UniProtKB: Guanine nucleotide-binding protein G(s) subunit alpha isoforms short |
-Macromolecule #3: FORSKOLIN
Macromolecule | Name: FORSKOLIN / type: ligand / ID: 3 / Number of copies: 1 / Formula: FOK |
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Molecular weight | Theoretical: 410.501 Da |
Chemical component information | ![]() ChemComp-FOK: |
-Macromolecule #4: 5'-GUANOSINE-DIPHOSPHATE-MONOTHIOPHOSPHATE
Macromolecule | Name: 5'-GUANOSINE-DIPHOSPHATE-MONOTHIOPHOSPHATE / type: ligand / ID: 4 / Number of copies: 1 / Formula: GSP |
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Molecular weight | Theoretical: 539.246 Da |
Chemical component information | ![]() ChemComp-GSP: |
-Macromolecule #5: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 5 / Number of copies: 1 / Formula: MG |
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Molecular weight | Theoretical: 24.305 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 5 mg/mL |
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Buffer | pH: 8 |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 55.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 0.6 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |