4FML

Catalytic domain of VahC from Aeromonas hydrophila

Summary for 4FML

• Entry DOI: 10.2210/pdb4fml/pdb
• Related: 3NTS
• Descriptor: VsdC (2 entities in total)
• Functional Keywords: transferase
• Biological source: Aeromonas hydrophila
• Total number of polymer chains: 3
• Total formula weight: 74225.34

Authors

Ravulapalli, R.,Kimber, M.S.,Merrill, A.R. (deposition date: 2012-06-18, release date: 2012-09-19, Last modification date: 2024-02-28)

Primary citation

Shniffer, A.,Visschedyk, D.D.,Ravulapalli, R.,Suarez, G.,Turgeon, Z.J.,Petrie, A.A.,Chopra, A.K.,Merrill, A.R.
Characterization of an actin-targeting ADP-ribosyltransferase from Aeromonas hydrophila.
J.Biol.Chem., 287:37030-37041, 2012

PubMed Abstract: The mono-ADP-ribosyltransferase (mART) toxins are contributing factors to a number of human diseases, including cholera, diphtheria, traveler's diarrhea, and whooping cough. VahC is a cytotoxic, actin-targeting mART from Aeromonas hydrophila PPD134/91. This bacterium is implicated primarily in diseases among freshwater fish species but also contributes to gastrointestinal and extraintestinal infections in humans. VahC was shown to ADP-ribosylate Arg-177 of actin, and the kinetic parameters were K(m)(NAD(+)) = 6 μM, K(m)(actin) = 24 μM, and k(cat) = 22 s(-1). VahC activity caused depolymerization of actin filaments, which induced caspase-mediated apoptosis in HeLa Tet-Off cells. Alanine-scanning mutagenesis of predicted catalytic residues showed the predicted loss of in vitro mART activity and cytotoxicity. Bioinformatic and kinetic analysis also identified three residues in the active site loop that were critical for the catalytic mechanism. A 1.9 Å crystal structure supported the proposed roles of these residues and their conserved nature among toxin homologues. Several small molecules were characterized as inhibitors of in vitro VahC mART activity and suramin was the best inhibitor (IC(50) = 20 μM). Inhibitor activity was also characterized against two other actin-targeting mART toxins. Notably, these inhibitors represent the first report of broad spectrum inhibition of actin-targeting mART toxins.

PubMed: 22969084
DOI: 10.1074/jbc.M112.397612

Experimental method

X-RAY DIFFRACTION (1.93 Å)