title: Structural and biophysical methods to analyze clock function and mechanism.
abstract: Structural approaches have provided insight into the mechanisms of circadian clock oscillators. This review focuses upon the myriad structural methods that have been applied to the molecular architecture of cyanobacterial circadian proteins, their interactions with each other, and the mechanism of the KaiABC posttranslational oscillator. X-ray crystallography and solution NMR were deployed to gain an understanding of the three-dimensional structures of the three proteins KaiA, KaiB, and KaiC that make up the inner timer in cyanobacteria. A hybrid structural biology approach including crystallography, electron microscopy, and solution scattering has shed light on the shapes of binary and ternary Kai protein complexes. Structural studies of the cyanobacterial oscillator demonstrate both the strengths and the limitations of the divide-and-conquer strategy. Thus, investigations of complexes involving domains and/or peptides have afforded valuable information into Kai protein interactions. However, high-resolution structural data are still needed at the level of complexes between the 360-kDa KaiC hexamer that forms the heart of the clock and its KaiA and KaiB partners.
authors: Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, Tennessee, USA. Electronic address: martin.egli@vanderbilt.edu.
authors: Martin
authors: M
authors: Egli
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000502:physiology; D056925:Circadian Rhythm Signaling Peptides and Proteins / Q000737:chemistry* / Q000502:physiology; D020285:Cryoelectron Microscopy; D018360:Crystallography, X-Ray; D000458:Cyanobacteria; D031541:Fluorescence Resonance Energy Transfer; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D053838:Scattering, Small Angle
chemical_list: D001426:Bacterial Proteins; D056925:Circadian Rhythm Signaling Peptides and Proteins; C114269:KaiA protein, cyanobacteria; C114270:KaiB protein, cyanobacteria; C114271:KaiC protein, cyanobacteria
keywords: ATPase; Circadian clock; Cyanobacteria; Hybrid approach; In vitro oscillator; KaiABC; Kinase; Phosphorylation; Protein–protein interactions; Structural biology