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- EMDB-22458: Structure of the Visual Signaling Complex between Transducin and ... -

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Basic information

Entry
Database: EMDB / ID: EMD-22458
TitleStructure of the Visual Signaling Complex between Transducin and Phosphodiesterase 6
Map dataVisual Signaling Complex between Transducin and Phosphodiesterase 6
Sample
  • Complex: Complex of Transducin and Phosphodiesterase 6
    • Complex: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha (E.C.3.1.4.35), Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta (E.C.3.1.4.35), Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiesterase subunit gamma (E.C.3.1.4.35)
      • Protein or peptide: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha
      • Protein or peptide: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta
      • Protein or peptide: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine nucleotide-binding protein G(i) subunit alpha-1 chimera
    • Complex: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine nucleotide-binding protein G(i) subunit alpha-1 chimera,Gt-alpha/Gi1-alpha chimera
      • Protein or peptide: Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiesterase subunit gamma
  • Ligand: ZINC ION
  • Ligand: MAGNESIUM ION
  • Ligand: 2-{2-ETHOXY-5-[(4-ETHYLPIPERAZIN-1-YL)SULFONYL]PHENYL}-5-METHYL-7-PROPYLIMIDAZO[5,1-F][1,2,4]TRIAZIN-4(1H)-ONE
  • Ligand: GUANOSINE-3',5'-MONOPHOSPHATE
  • Ligand: GUANOSINE-5'-TRIPHOSPHATE
KeywordsG protein / G protein-effector complex / Transducin / phosphodiesterase 6 / phototransduction / GPCR signaling / SIGNALING PROTEIN
Function / homology
Function and homology information


detection of light stimulus involved in visual perception / negative regulation of cyclic-nucleotide phosphodiesterase activity / 3',5'-cyclic-GMP phosphodiesterase / G protein-coupled receptor complex / Inactivation, recovery and regulation of the phototransduction cascade / positive regulation of G protein-coupled receptor signaling pathway / phototransduction, visible light / Activation of the phototransduction cascade / Ca2+ pathway / positive regulation of epidermal growth factor receptor signaling pathway ...detection of light stimulus involved in visual perception / negative regulation of cyclic-nucleotide phosphodiesterase activity / 3',5'-cyclic-GMP phosphodiesterase / G protein-coupled receptor complex / Inactivation, recovery and regulation of the phototransduction cascade / positive regulation of G protein-coupled receptor signaling pathway / phototransduction, visible light / Activation of the phototransduction cascade / Ca2+ pathway / positive regulation of epidermal growth factor receptor signaling pathway / photoreceptor outer segment membrane / G alpha (i) signalling events / entrainment of circadian clock by photoperiod / phototransduction / cGMP binding / 3',5'-cyclic-GMP phosphodiesterase activity / response to light stimulus / acyl binding / 3',5'-cyclic-AMP phosphodiesterase activity / photoreceptor inner segment / cAMP-mediated signaling / visual perception / G protein-coupled receptor binding / G-protein beta/gamma-subunit complex binding / adenylate cyclase-modulating G protein-coupled receptor signaling pathway / photoreceptor disc membrane / GDP binding / heterotrimeric G-protein complex / retina development in camera-type eye / molecular adaptor activity / GTPase activity / GTP binding / protein kinase binding / zinc ion binding / metal ion binding / cytoplasm
Similarity search - Function
Retinal cGMP phosphodiesterase, gamma subunit / Retinal cGMP phosphodiesterase, gamma subunit superfamily / Retinal cGMP phosphodiesterase, gamma subunit / 3'5'-cyclic nucleotide phosphodiesterase / 3'5'-cyclic nucleotide phosphodiesterase, catalytic domain / 3'5'-cyclic nucleotide phosphodiesterase, conserved site / 3'5'-cyclic nucleotide phosphodiesterase, catalytic domain superfamily / 3'5'-cyclic nucleotide phosphodiesterase / 3'5'-cyclic nucleotide phosphodiesterase domain signature. / 3'5'-cyclic nucleotide phosphodiesterase domain profile. ...Retinal cGMP phosphodiesterase, gamma subunit / Retinal cGMP phosphodiesterase, gamma subunit superfamily / Retinal cGMP phosphodiesterase, gamma subunit / 3'5'-cyclic nucleotide phosphodiesterase / 3'5'-cyclic nucleotide phosphodiesterase, catalytic domain / 3'5'-cyclic nucleotide phosphodiesterase, conserved site / 3'5'-cyclic nucleotide phosphodiesterase, catalytic domain superfamily / 3'5'-cyclic nucleotide phosphodiesterase / 3'5'-cyclic nucleotide phosphodiesterase domain signature. / 3'5'-cyclic nucleotide phosphodiesterase domain profile. / GAF domain / Domain present in phytochromes and cGMP-specific phosphodiesterases. / GAF domain / GAF-like domain superfamily / Metal dependent phosphohydrolases with conserved 'HD' motif. / HD/PDEase domain / G-protein alpha subunit, group I / Guanine nucleotide binding protein (G-protein), alpha subunit / G protein alpha subunit, helical insertion / G-protein alpha subunit / G-alpha domain profile. / G protein alpha subunit / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
Guanine nucleotide-binding protein G(t) subunit alpha-1 / Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiesterase subunit gamma / Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha / Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta
Similarity search - Component
Biological speciesBos taurus (cattle)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.2 Å
AuthorsGao Y / Eskici G
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R35 GM122575 United States
National Institutes of Health/National Cancer Institute (NIH/NCI)R01 CA201402 United States
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)R01 NS092695 United States
CitationJournal: Mol Cell / Year: 2020
Title: Structure of the Visual Signaling Complex between Transducin and Phosphodiesterase 6.
Authors: Yang Gao / Gözde Eskici / Sekar Ramachandran / Frédéric Poitevin / Alpay Burak Seven / Ouliana Panova / Georgios Skiniotis / Richard A Cerione /
Abstract: Heterotrimeric G proteins communicate signals from activated G protein-coupled receptors to downstream effector proteins. In the phototransduction pathway responsible for vertebrate vision, the G ...Heterotrimeric G proteins communicate signals from activated G protein-coupled receptors to downstream effector proteins. In the phototransduction pathway responsible for vertebrate vision, the G protein-effector complex is composed of the GTP-bound transducin α subunit (Gα·GTP) and the cyclic GMP (cGMP) phosphodiesterase 6 (PDE6), which stimulates cGMP hydrolysis, leading to hyperpolarization of the photoreceptor cell. Here we report a cryo-electron microscopy (cryoEM) structure of PDE6 complexed to GTP-bound Gα. The structure reveals two Gα·GTP subunits engaging the PDE6 hetero-tetramer at both the PDE6 catalytic core and the PDEγ subunits, driving extensive rearrangements to relieve all inhibitory constraints on enzyme catalysis. Analysis of the conformational ensemble in the cryoEM data highlights the dynamic nature of the contacts between the two Gα·GTP subunits and PDE6 that supports an alternating-site catalytic mechanism.
History
DepositionAug 15, 2020-
Header (metadata) releaseOct 21, 2020-
Map releaseOct 21, 2020-
UpdateMay 29, 2024-
Current statusMay 29, 2024Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.007
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 0.007
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  • Surface view with fitted model
  • Atomic models: PDB-7jsn
  • Surface level: 0.007
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_22458.png

Downloads & links

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Map

FileDownload / File: emd_22458.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationVisual Signaling Complex between Transducin and Phosphodiesterase 6
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.85 Å/pix.
x 320 pix.
= 272. Å		0.85 Å/pix.
x 320 pix.
= 272. Å		0.85 Å/pix.
x 320 pix.
= 272. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.85 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.007 / Movie #1: 0.007
Minimum - Maximum-0.002930495 - 0.059562024
Average (Standard dev.)0.000012298156 (±0.0018455955)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions320320320
Spacing320320320
CellA=B=C: 272.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.850.850.85
M x/y/z320320320
origin x/y/z0.0000.0000.000
length x/y/z272.000272.000272.000
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ304304304
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS320320320
D min/max/mean-0.0030.0600.000

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Supplemental data

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Sample components

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Entire : Complex of Transducin and Phosphodiesterase 6

EntireName: Complex of Transducin and Phosphodiesterase 6
Components
  • Complex: Complex of Transducin and Phosphodiesterase 6
    • Complex: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha (E.C.3.1.4.35), Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta (E.C.3.1.4.35), Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiesterase subunit gamma (E.C.3.1.4.35)
      • Protein or peptide: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha
      • Protein or peptide: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta
      • Protein or peptide: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine nucleotide-binding protein G(i) subunit alpha-1 chimera
    • Complex: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine nucleotide-binding protein G(i) subunit alpha-1 chimera,Gt-alpha/Gi1-alpha chimera
      • Protein or peptide: Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiesterase subunit gamma
  • Ligand: ZINC ION
  • Ligand: MAGNESIUM ION
  • Ligand: 2-{2-ETHOXY-5-[(4-ETHYLPIPERAZIN-1-YL)SULFONYL]PHENYL}-5-METHYL-7-PROPYLIMIDAZO[5,1-F][1,2,4]TRIAZIN-4(1H)-ONE
  • Ligand: GUANOSINE-3',5'-MONOPHOSPHATE
  • Ligand: GUANOSINE-5'-TRIPHOSPHATE

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Supramolecule #1: Complex of Transducin and Phosphodiesterase 6

SupramoleculeName: Complex of Transducin and Phosphodiesterase 6 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#4
Molecular weightTheoretical: 300 KDa

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Supramolecule #2: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha (E...

SupramoleculeName: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha (E.C.3.1.4.35), Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta (E.C.3.1.4.35), Retinal rod rhodopsin-sensitive cGMP ...Name: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha (E.C.3.1.4.35), Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta (E.C.3.1.4.35), Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiesterase subunit gamma (E.C.3.1.4.35)
type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#3
Source (natural)Organism: Bos taurus (cattle)

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Supramolecule #3: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine ...

SupramoleculeName: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine nucleotide-binding protein G(i) subunit alpha-1 chimera,Gt-alpha/Gi1-alpha chimera
type: complex / ID: 3 / Parent: 1 / Macromolecule list: #4
Source (natural)Organism: Bos taurus (cattle)

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Macromolecule #1: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha

MacromoleculeName: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha
type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: 3',5'-cyclic-GMP phosphodiesterase
Source (natural)Organism: Bos taurus (cattle)
Molecular weightTheoretical: 99.461789 KDa
SequenceString: MGEVTAEEVE KFLDSNVSFA KQYYNLRYRA KVISDLLGPR EAAVDFSNYH ALNSVEESEI IFDLLRDFQD NLQAEKCVFN VMKKLCFLL QADRMSLFMY RARNGIAELA TRLFNVHKDA VLEECLVAPD SEIVFPLDMG VVGHVALSKK IVNVPNTEED E HFCDFVDT ...String:
MGEVTAEEVE KFLDSNVSFA KQYYNLRYRA KVISDLLGPR EAAVDFSNYH ALNSVEESEI IFDLLRDFQD NLQAEKCVFN VMKKLCFLL QADRMSLFMY RARNGIAELA TRLFNVHKDA VLEECLVAPD SEIVFPLDMG VVGHVALSKK IVNVPNTEED E HFCDFVDT LTEYQTKNIL ASPIMNGKDV VAIIMVVNKV DGPHFTENDE EILLKYLNFA NLIMKVFHLS YLHNCETRRG QI LLWSGSK VFEELTDIER QFHKALYTVR AFLNCDRYSV GLLDMTKQKE FFDVWPVLMG EAPPYAGPRT PDGREINFYK VID YILHGK EDIKVIPNPP PDHWALVSGL PTYVAQNGLI CNIMNAPSED FFAFQKEPLD ESGWMIKNVL SMPIVNKKEE IVGV ATFYN RKDGKPFDEM DETLMESLTQ FLGWSVLNPD TYELMNKLEN RKDIFQDMVK YHVKCDNEEI QTILKTREVY GKEPW ECEE EELAEILQGE LPDADKYEIN KFHFSDLPLT ELELVKCGIQ MYYELKVVDK FHIPQEALVR FMYSLSKGYR RITYHN WRH GFNVGQTMFS LLVTGKLKRY FTDLEALAMV TAAFCHDIDH RGTNNLYQMK SQNPLAKLHG SSILERHHLE FGKTLLR DE SLNIFQNLNR RQHEHAIHMM DIAIIATDLA LYFKKRTMFQ KIVDQSKTYE TQQEWTQYMM LDQTRKEIVM AMMMTACD L SAITKPWEVQ SKVALLVAAE FWEQGDLERT VLQQNPIPMM DRNKADELPK LQVGFIDFVC TFVYKEFSRF HEEITPMLD GITNNRKEWK ALADEYETKM KGLEEEKQKQ QAANQAAAGS QHGGKQPGGG PASKSCCVQ

UniProtKB: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit alpha

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Macromolecule #2: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta

MacromoleculeName: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta
type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: 3',5'-cyclic-GMP phosphodiesterase
Source (natural)Organism: Bos taurus (cattle)
Molecular weightTheoretical: 98.449648 KDa
SequenceString: MSLSEGQVHR FLDQNPGFAD QYFGRKLSPE DVANACEDGC PEGCTSFREL CQVEESAALF ELVQDMQENV NMERVVFKIL RRLCSILHA DRCSLFMYRQ RNGVAELATR LFSVQPDSVL EDCLVPPDSE IVFPLDIGVV GHVAQTKKMV NVQDVMECPH F SSFADELT ...String:
MSLSEGQVHR FLDQNPGFAD QYFGRKLSPE DVANACEDGC PEGCTSFREL CQVEESAALF ELVQDMQENV NMERVVFKIL RRLCSILHA DRCSLFMYRQ RNGVAELATR LFSVQPDSVL EDCLVPPDSE IVFPLDIGVV GHVAQTKKMV NVQDVMECPH F SSFADELT DYVTRNILAT PIMNGKDVVA VIMAVNKLDG PCFTSEDEDV FLKYLNFGTL NLKIYHLSYL HNCETRRGQV LL WSANKVF EELTDIERQF HKAFYTVRAY LNCDRYSVGL LDMTKEKEFF DVWPVLMGEA QAYSGPRTPD GREILFYKVI DYI LHGKED IKVIPSPPAD HWALASGLPT YVAESGFICN IMNAPADEMF NFQEGPLDDS GWIVKNVLSM PIVNKKEEIV GVAT FYNRK DGKPFDEQDE VLMESLTQFL GWSVLNTDTY DKMNKLENRK DIAQDMVLYH VRCDREEIQL ILPTRERLGK EPADC EEDE LGKILKEVLP GPAKFDIYEF HFSDLECTEL ELVKCGIQMY YELGVVRKFQ IPQEVLVRFL FSVSKGYRRI TYHNWR HGF NVAQTMFTLL MTGKLKSYYT DLEAFAMVTA GLCHDIDHRG TNNLYQMKSQ NPLAKLHGSS ILERHHLEFG KFLLSEE TL NIYQNLNRRQ HEHVIHLMDI AIIATDLALY FKKRTMFQKI VDESKNYEDR KSWVEYLSLE TTRKEIVMAM MMTACDLS A ITKPWEVQSK VALLVAAEFW EQGDLERTVL DQQPIPMMDR NKAAELPKLQ VGFIDFVCTF VYKEFSRFHE EILPMFDRL QNNRKEWKAL ADEYEAKVKA LEEDQKKETT AKKVGTEICN GGPAPRSSTC RIL

UniProtKB: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta

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Macromolecule #3: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine ...

MacromoleculeName: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine nucleotide-binding protein G(i) subunit alpha-1 chimera
type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Bos taurus (cattle)
Molecular weightTheoretical: 42.218035 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MAHHHHHHAM GAGASAEEKH SRELEKKLKE DAEKDARTVK LLLLGAGESG KSTIVKQMKI IHQDGYSLEE CLEFIAIIYG NTLQSILAI VRAMTTLNIQ YGDSARQDDA RKLMHMADTI EEGTMPKEMS DIIQRLWKDS GIQACFDRAS EYQLNDSAGY Y LSDLERLV ...String:
MAHHHHHHAM GAGASAEEKH SRELEKKLKE DAEKDARTVK LLLLGAGESG KSTIVKQMKI IHQDGYSLEE CLEFIAIIYG NTLQSILAI VRAMTTLNIQ YGDSARQDDA RKLMHMADTI EEGTMPKEMS DIIQRLWKDS GIQACFDRAS EYQLNDSAGY Y LSDLERLV TPGYVPTEQD VLRSCVKTTG IIETQFSFKD LNFRMFDVGG LRSERKKWIH CFEGVTAIIF CVALSDYDMV LV EDDEVNR MHESMHLFNS ICNNKWFTDT SIILFLNKKD LFEEKIKKSP LSICFPDYAG SNTYEEAGNY IKVQFLELNM RRD VKEIYS HMTCATDTQN VKFVFDAVTD IIIKENLKDC GLFAAATETS QVAPA

UniProtKB: Guanine nucleotide-binding protein G(t) subunit alpha-1, Guanine nucleotide-binding protein G(t) subunit alpha-1

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Macromolecule #4: Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiestera...

MacromoleculeName: Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiesterase subunit gamma
type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO / EC number: 3',5'-cyclic-GMP phosphodiesterase
Source (natural)Organism: Bos taurus (cattle)
Molecular weightTheoretical: 9.684229 KDa
SequenceString:
MNLEPPKAEI RSATRVMGGP VTPRKGPPKF KQRQTRQFKS KPPKKGVQGF GDDIPGMEGL GTDITVICPW EAFNHLELHE LAQYGII

UniProtKB: Retinal rod rhodopsin-sensitive cGMP 3',5'-cyclic phosphodiesterase subunit gamma

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Macromolecule #5: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 5 / Number of copies: 2 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

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Macromolecule #6: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 6 / Number of copies: 2 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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Macromolecule #7: 2-{2-ETHOXY-5-[(4-ETHYLPIPERAZIN-1-YL)SULFONYL]PHENYL}-5-METHYL-7...

MacromoleculeName: 2-{2-ETHOXY-5-[(4-ETHYLPIPERAZIN-1-YL)SULFONYL]PHENYL}-5-METHYL-7-PROPYLIMIDAZO[5,1-F][1,2,4]TRIAZIN-4(1H)-ONE
type: ligand / ID: 7 / Number of copies: 2 / Formula: VDN
Molecular weightTheoretical: 488.603 Da
Chemical component information

ChemComp-VDN:
2-{2-ETHOXY-5-[(4-ETHYLPIPERAZIN-1-YL)SULFONYL]PHENYL}-5-METHYL-7-PROPYLIMIDAZO[5,1-F][1,2,4]TRIAZIN-4(1H)-ONE / medication, inhibitor*YM

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Macromolecule #8: GUANOSINE-3',5'-MONOPHOSPHATE

MacromoleculeName: GUANOSINE-3',5'-MONOPHOSPHATE / type: ligand / ID: 8 / Number of copies: 2 / Formula: 35G
Molecular weightTheoretical: 345.205 Da
Chemical component information

ChemComp-35G:
GUANOSINE-3',5'-MONOPHOSPHATE

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Macromolecule #9: GUANOSINE-5'-TRIPHOSPHATE

MacromoleculeName: GUANOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 9 / Number of copies: 2 / Formula: GTP
Molecular weightTheoretical: 523.18 Da
Chemical component information

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration2 mg/mL
BufferpH: 8 / Details: 20 mM Tris pH 8.0, 5 mM MgCl2 and 1 uM vardenafil
GridModel: Quantifoil R1.2/1.3 / Material: GOLD / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 295 K / Instrument: FEI VITROBOT MARK IV
Details0.05% octyl glucoside was used as an additive.

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average exposure time: 4.0 sec. / Average electron dose: 48.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: EMDB MAP
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 143125
Initial angle assignmentType: PROJECTION MATCHING
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementProtocol: BACKBONE TRACE
Output model

PDB-7jsn:
Structure of the Visual Signaling Complex between Transducin and Phosphodiesterase 6

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