Unknown

Dataset Information

0

Proximity-induced caspase-9 activation on a DNA origami-based synthetic apoptosome.


ABSTRACT: Living cells regulate key cellular processes by spatial organisation of catalytically active proteins in higher-order signalling complexes. These act as organising centres to facilitate proximity-induced activation and inhibition of multiple intrinsically weakly associating signalling components, which makes elucidation of the underlying protein-protein interactions challenging. Here we show that DNA origami nanostructures provide a programmable molecular platform for the systematic analysis of signalling proteins by engineering a synthetic DNA origami-based version of the apoptosome, a multi-protein complex that regulates apoptosis by co-localizing multiple caspase-9 monomers. Tethering of both wildtype and inactive caspase-9 variants to a DNA origami platform demonstrates that enzymatic activity is induced by proximity-driven dimerization with half-of-sites reactivity, and additionally, reveals a multivalent activity enhancement in oligomers of three and four enzymes. Our results offer fundamental insights in caspase-9 activity regulation and demonstrate that DNA origami-based protein assembly platforms have the potential to inform the function of other multi-enzyme complexes involved in inflammation, innate immunity and cell death.

SUBMITTER: Rosier BJHM 

PROVIDER: S-EPMC7080557 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

altmetric image

Publications

Proximity-induced caspase-9 activation on a DNA origami-based synthetic apoptosome.

Rosier Bas J H M BJHM   Markvoort Albert J AJ   Gumí Audenis Berta B   Roodhuizen Job A L JAL   den Hamer Anniek A   Brunsveld Luc L   de Greef Tom F A TFA  

Nature catalysis 20200106 3


Living cells regulate key cellular processes by spatial organisation of catalytically active proteins in higher-order signalling complexes. These act as organising centres to facilitate proximity-induced activation and inhibition of multiple intrinsically weakly associating signalling components, which makes elucidation of the underlying protein-protein interactions challenging. Here we show that DNA origami nanostructures provide a programmable molecular platform for the systematic analysis of  ...[more]

Similar Datasets

| S-EPMC2755603 | biostudies-literature
| S-EPMC3100593 | biostudies-literature
| S-EPMC4246342 | biostudies-literature
| S-EPMC7198528 | biostudies-literature
| S-EPMC1636747 | biostudies-literature
| S-EPMC3155825 | biostudies-literature
| S-EPMC10416349 | biostudies-literature
| S-EPMC5320974 | biostudies-literature
| S-EPMC4318144 | biostudies-literature
| S-EPMC3245238 | biostudies-literature