Journal
MOLECULAR MICROBIOLOGY
Volume 94, Issue 4, Pages 898-912Publisher
WILEY
DOI: 10.1111/mmi.12807
Keywords
-
Categories
Funding
- NIH [R01 AI073783, R01 AI099493, R01 AI031940]
Ask authors/readers for more resources
The catalytic A1 subunit of cholera toxin (CTA1) has a disordered structure at 37 degrees C. An interaction with host factors must therefore place CTA1 in a folded conformation for the modification of its Gs target which resides in a lipid raft environment. Host ADP-ribosylation factors (ARFs) act as in vitro allosteric activators of CTA1, but the molecular events of this process are not fully characterized. Isotope-edited Fourier transform infrared spectroscopy monitored ARF6-induced structural changes to CTA1, which were correlated to changes in CTA1 activity. We found ARF6 prevents the thermal disordering of structured CTA1 and stimulates the activity of stabilized CTA1 over a range of temperatures. Yet ARF6 alone did not promote the refolding of disordered CTA1 to an active state. Instead, lipid rafts shifted disordered CTA1 to a folded conformation with a basal level of activity that could be further stimulated by ARF6. Thus, ARF alone is unable to activate disordered CTA1 at physiological temperature: additional host factors such as lipid rafts place CTA1 in the folded conformation required for its ARF-mediated activation. Interaction with ARF is required for in vivo toxin activity, as enzymatically active CTA1 mutants that cannot be further stimulated by ARF6 fail to intoxicate cultured cells.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available