Adenylate cyclase toxin from Bordetella pertussis. Conformational change associated with toxin activity.
Adenylate cyclase (AC) toxin from Bordetella pertussis interacts with and enters eukaryotic cells to catalyze the production of supraphysiologic levels of cyclic AMP. Although the calmodulin-activated enzymatic activity (ability to convert ATP to cyclic AMP in a cell-free assay) of this molecule is calcium independent, its toxin activity (ability to increase cyclic AMP levels in intact target cells) requires extracellular calcium. Toxin activity as a function of calcium concentration is biphasic, with no intoxication occurring in the absence of calcium, low level intoxication (200-300 pmol of cyclic AMP/mg of Jurkat cell protein) occurring with free calcium concentrations between 100 nM and 100 microM and a 10-fold increase in AC toxin activity at free calcium concentrations above 300 microM. The molecule exhibits a conformational change when free calcium concentrations exceed 100 microM as demonstrated by shift in intrinsic tryptophan fluorescence, an alteration in binding of one anti-AC monoclonal antibody, protection of a fragment from trypsin-mediated proteolysis, and a structural modification as illustrated by electron microscopy. Thus, it appears that an increase in the ambient calcium concentration to a critical point and the ensuing interaction of the toxin with calcium induces a conformational change which is necessary for its insertion into the target cell and for delivery of its catalytic domain to the cell interior.
Hewlett
E L
EL
Department of Medicine, University of Virginia School of Medicine, Charlottesville 22908.
Gray
L
L
Allietta
M
M
Ehrmann
I
I
Gordon
V M
VM
Gray
M C
MC
D037361:Adenylate Cyclase Toxin; D000494:Allosteric Regulation; D001886:Bordetella pertussis / Q000378:metabolism*; D002118:Calcium / Q000502:physiology*; D005453:Fluorescence; D008854:Microscopy, Electron; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D014357:Trypsin; D010566:Virulence Factors, Bordetella / Q000737:chemistry / Q000378:metabolism*
D037361:Adenylate Cyclase Toxin; D010566:Virulence Factors, Bordetella; D014357:Trypsin; D002118:Calcium