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The Adenylate Cyclase (CyaA) Toxin from Bordetella pertussis Has No Detectable Phospholipase A (PLA) Activity In Vitro

1
Chemistry and Structural Biology Department, Institut Pasteur, UMR CNRS 3528, CEDEX 15, 75724 Paris, France
2
Université Paris Diderot Paris VII, 75013 Paris, France
3
Mass Spectrometry for Biology Unit, Proteomics Platform, Institut Pasteur, USR CNRS 2000, CEDEX 15, 75724 Paris, France
*
Authors to whom correspondence should be addressed.
Toxins 2019, 11(2), 111; https://doi.org/10.3390/toxins11020111
Received: 28 January 2019 / Accepted: 11 February 2019 / Published: 13 February 2019
(This article belongs to the Special Issue RTX Toxins)
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Abstract

The adenylate cyclase (CyaA) toxin produced in Bordetella pertussis is the causative agent of whooping cough. CyaA exhibits the remarkable capacity to translocate its N-terminal adenyl cyclase domain (ACD) directly across the plasma membrane into the cytosol of eukaryotic cells. Once translocated, calmodulin binds and activates ACD, leading to a burst of cAMP that intoxicates the target cell. Previously, Gonzalez-Bullon et al. reported that CyaA exhibits a phospholipase A activity that could destabilize the membrane to facilitate ACD membrane translocation. However, Bumba and collaborators lately reported that they could not replicate these results. To clarify this controversy, we assayed the putative PLA activity of two CyaA samples purified in two different laboratories by using two distinct fluorescent probes reporting either PLA2 or both PLA1 and PLA2 activities, as well as in various experimental conditions (i.e., neutral or negatively charged membranes in different buffers.) However, we could not detect any PLA activity in these CyaA batches. Thus, our data independently confirm that CyaA does not possess any PLA activity. View Full-Text
Keywords: adenylate cyclase toxin; CyaA toxin; bordetella pertussis; phospholipase A; fluorescence adenylate cyclase toxin; CyaA toxin; bordetella pertussis; phospholipase A; fluorescence
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Voegele, A.; Sadi, M.; Raoux-Barbot, D.; Douché, T.; Matondo, M.; Ladant, D.; Chenal, A. The Adenylate Cyclase (CyaA) Toxin from Bordetella pertussis Has No Detectable Phospholipase A (PLA) Activity In Vitro. Toxins 2019, 11, 111.

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