Solution NMR spectroscopy of supra-molecular systems, why bother? A methyl-TROSY view

Kay, Lewis E.

doi:10.1016/j.jmr.2011.03.008

Cited by 71 publications

(61 citation statements)

References 61 publications

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“…The molecular details of KaiC-KaiB binding remain incompletely understood, however. Formation of this supramolecular complex (20) can be monitored by NMR spectroscopy (25)(26)(27) (Fig. 1A, column 1) on the addition of S431E-KaiC, a phosphomimic of pST-KaiC (Fig.…”

Section: Resultsmentioning

confidence: 99%

Rhythmic ring–ring stacking drives the circadian oscillator clockwise

Chang¹,

Tseng

Kuo³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

100

163

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The oscillator of the circadian clock of cyanobacteria is composed of three proteins, KaiA, KaiB, and KaiC, which together generate a selfsustained ∼24-h rhythm of phosphorylation of KaiC. The mechanism propelling this oscillator has remained elusive, however. We show that stacking interactions between the CI and CII rings of KaiC drive the transition from the phosphorylation-specific KaiC-KaiA interaction to the dephosphorylation-specific KaiC-KaiB interaction. We have identified the KaiB-binding site, which is on the CI domain. This site is hidden when CI domains are associated as a hexameric ring. However, stacking of the CI and CII rings exposes the KaiB-binding site. Because the clock output protein SasA also binds to CI and competes with KaiB for binding, ring stacking likely regulates clock output. We demonstrate that ADP can expose the KaiB-binding site in the absence of ring stacking, providing an explanation for how it can reset the clock.dynamic allostery | NMR | protein structure | kinase | kinetics

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Section: Resultsmentioning

confidence: 99%

Rhythmic ring–ring stacking drives the circadian oscillator clockwise

Chang¹,

Tseng

Kuo³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

100

163

View full text Add to dashboard Cite

show abstract

“…In addition, methyl-TROSY based experiments have guided new approaches to developing proteasome inhibitors by demonstrating that inhibition can be achieved by binding in the vicinity of the interface between α and β subunits in a manner independent of binding to the active sites [251]. Overall, these remarkable achievements were made possible by the combination of selective labeling of methyl groups of amino acids in a deuterated background and the development of methyl-TROSY [6].…”

Section: Proteasomementioning

confidence: 99%

“…On the other hand, this flexibility makes such complexes suitable targets for NMR spectroscopy, which in recent years became applicable to large protein sizes through advanced techniques such as specialized labeling schemes and methyl TROSY spectroscopy [5][6][7].…”

Section: Introductionmentioning

confidence: 99%

Chaperones and chaperone–substrate complexes: Dynamic playgrounds for NMR spectroscopists

Burmann¹,

Hiller²

2015

Progress in Nuclear Magnetic Resonance Spectroscopy

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“…Recent advances in NMR spectroscopy (28,29) allow its application to supramolecular protein particles (30,31). KaiC, with a mass of 345 kDa, belongs to the supramolecular class.…”

mentioning

confidence: 99%

Flexibility of the C-terminal, or CII, ring of KaiC governs the rhythm of the circadian clock of cyanobacteria

Chang

Kuo

Tseng

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

144

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In the cyanobacterial circadian oscillator, KaiA and KaiB alternately stimulate autophosphorylation and autodephosphorylation of KaiC with a periodicity of approximately 24 h. KaiA activates autophosphorylation by selectively capturing the A loops of KaiC in their exposed positions. The A loops and sites of phosphorylation, residues S431 and T432, are located in the CII ring of KaiC. We find that the flexibility of the CII ring governs the rhythm of KaiC autophosphorylation and autodephosphorylation and is an example of dynamics-driven protein allostery. KaiA-induced autophosphorylation requires flexibility of the CII ring. In contrast, rigidity is required for KaiC-KaiB binding, which induces a conformational change in KaiB that enables it to sequester KaiA by binding to KaiA's linker. Autophosphorylation of the S431 residues around the CII ring stabilizes the CII ring, making it rigid. In contrast, autophosphorylation of the T432 residues offsets phospho-S431-induced rigidity to some extent. In the presence of KaiA and KaiB, the dynamic states of the CII ring of KaiC executes the following circadian rhythm: CII ST flexible → CII SpT flexible → CII pSpT rigid → CII pST very-rigid → CII ST flexible . Apparently, these dynamic states govern the pattern of phosphorylation, ST → SpT → pSpT → pST → ST. CII-CI ring-on-ring stacking is observed when the CII ring is rigid, suggesting a mechanism through which the ATPase activity of the CI ring is rhythmically controlled. SasA, a circadian clock-output protein, binds to the CI ring. Thus, rhythmic ring stacking may also control clock-output pathways.

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Solution NMR spectroscopy of supra-molecular systems, why bother? A methyl-TROSY view

Cited by 71 publications

References 61 publications

Rhythmic ring–ring stacking drives the circadian oscillator clockwise

Rhythmic ring–ring stacking drives the circadian oscillator clockwise

Chaperones and chaperone–substrate complexes: Dynamic playgrounds for NMR spectroscopists

Flexibility of the C-terminal, or CII, ring of KaiC governs the rhythm of the circadian clock of cyanobacteria

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