|Entry||Database: EMDB / ID: 3603|
|Title||KaiCB circadian clock backbone model based on a Cryo-EM density|
|Map data||Circadian Clock Complex KaiBC|
|Sample||KaiCB circadian clock complex|
|Function/homology||Circadian clock protein KaiB / KaiB domain / regulation of phosphorelay signal transduction system / KaiB domain / Circadian clock protein kinase KaiC / Circadian clock KaiC, bacteria / KaiC domain / KaiC domain profile. / KaiC-like domain / entrainment of circadian clock ...Circadian clock protein KaiB / KaiB domain / regulation of phosphorelay signal transduction system / KaiB domain / Circadian clock protein kinase KaiC / Circadian clock KaiC, bacteria / KaiC domain / KaiC domain profile. / KaiC-like domain / entrainment of circadian clock / negative regulation of circadian rhythm / KaiC / negative regulation of phosphorylation / regulation of circadian rhythm / rhythmic process / circadian rhythm / Thioredoxin-like superfamily / non-specific serine/threonine protein kinase / protein serine/threonine kinase activity / magnesium ion binding / regulation of transcription, DNA-templated / P-loop containing nucleoside triphosphate hydrolase / transcription, DNA-templated / DNA binding / ATP binding / identical protein binding / plasma membrane / cytoplasm / Circadian clock protein kinase KaiC / Circadian clock protein KaiB|
Function and homology information
|Source||Synechococcus elongatus / bacteria / cyanobacteria /|
|Method||single particle reconstruction, at 7 Å resolution|
|Authors||Schuller JM / Snijder J / Loessl P / Heck AJR / Foerster F|
|Citation||Journal: Science / Year: 2017|
Title: Structures of the cyanobacterial circadian oscillator frozen in a fully assembled state.
Authors: Joost Snijder / Jan M Schuller / Anika Wiegard / Philip Lössl / Nicolas Schmelling / Ilka M Axmann / Jürgen M Plitzko / Friedrich Förster / Albert J R Heck
Abstract: Cyanobacteria have a robust circadian oscillator, known as the Kai system. Reconstituted from the purified protein components KaiC, KaiB, and KaiA, it can tick autonomously in the presence of ...Cyanobacteria have a robust circadian oscillator, known as the Kai system. Reconstituted from the purified protein components KaiC, KaiB, and KaiA, it can tick autonomously in the presence of adenosine 5'-triphosphate (ATP). The KaiC hexamers enter a natural 24-hour reaction cycle of autophosphorylation and assembly with KaiB and KaiA in numerous diverse forms. We describe the preparation of stoichiometrically well-defined assemblies of KaiCB and KaiCBA, as monitored by native mass spectrometry, allowing for a structural characterization by single-particle cryo-electron microscopy and mass spectrometry. Our data reveal details of the interactions between the Kai proteins and provide a structural basis to understand periodic assembly of the protein oscillator.
Copyright: 2017, American Association for the Advancement of Science.
|Date||Deposition: Feb 24, 2017 / Header (metadata) release: Mar 29, 2017 / Map release: Apr 12, 2017 / Last update: Jul 12, 2017|
Downloads & links
|File||emd_3603.map.gz (map file in CCP4 format, 30119 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 1.35 Å|
CCP4 map header:
-Entire KaiCB circadian clock complex
|Entire||Name: KaiCB circadian clock complex / Number of components: 1|
-Component #1: protein, KaiCB circadian clock complex
|Sample solution||pH: 7.5|
|Vitrification||Instrument: HOMEMADE PLUNGER / Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 45 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: FEI TITAN KRIOS AUTOGRID HOLDER|
|Camera||Detector: GATAN K2 (4k x 4k)|
|Processing||Method: single particle reconstruction / Number of projections: 5137|
|3D reconstruction||Resolution: 7 Å / Resolution method: FSC 0.143 CUT-OFF|
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