Calmodulin fishing with a structurally disordered bait triggers CyaA catalysis.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101183755 Publication Model: eCollection Cited Medium: Internet ISSN: 1545-7885 (Electronic) Linking ISSN: 15449173 NLM ISO Abbreviation: PLoS Biol Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science, [2003]-

Adenylate Cyclase Toxin/*chemistry ; Bordetella pertussis/*chemistry ; Calmodulin/*chemistry; Adenylate Cyclase Toxin/metabolism ; Adenylate Cyclase Toxin/physiology ; Bordetella pertussis/pathogenicity ; Calcium Signaling ; Calmodulin/metabolism ; Calmodulin/physiology ; Catalysis ; Catalytic Domain ; Circular Dichroism ; Cyclic AMP/metabolism ; Deuterium Exchange Measurement ; Mass Spectrometry ; Models, Molecular ; Protein Binding ; Protein Conformation ; Scattering, Small Angle ; Synchrotrons

Once translocated into the cytosol of target cells, the catalytic domain (AC) of the adenylate cyclase toxin (CyaA), a major virulence factor of Bordetella pertussis, is potently activated by binding calmodulin (CaM) to produce supraphysiological levels of cAMP, inducing cell death. Using a combination of small-angle X-ray scattering (SAXS), hydrogen/deuterium exchange mass spectrometry (HDX-MS), and synchrotron radiation circular dichroism (SR-CD), we show that, in the absence of CaM, AC exhibits significant structural disorder, and a 75-residue-long stretch within AC undergoes a disorder-to-order transition upon CaM binding. Beyond this local folding, CaM binding induces long-range allosteric effects that stabilize the distant catalytic site, whilst preserving catalytic loop flexibility. We propose that the high enzymatic activity of AC is due to a tight balance between the CaM-induced decrease of structural flexibility around the catalytic site and the preservation of catalytic loop flexibility, allowing for fast substrate binding and product release. The CaM-induced dampening of AC conformational disorder is likely relevant to other CaM-activated enzymes.

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0 (Adenylate Cyclase Toxin)
0 (Calmodulin)
E0399OZS9N (Cyclic AMP)

Date Created: 20171230 Date Completed: 20190128 Latest Revision: 20190128

20221213

PMC5764468

10.1371/journal.pbio.2004486

29287065