Implication of pH in the catalytic properties of anthrax lethal factor.
The anthrax lethal factor (LF) is a Zn(2+)-endopeptidase specific for mitogen-activated protein kinase kinases (MAPKKs), which are cleaved within their N-terminal region. Much line of effort was carried out to elucidate the catalytic activity of LF for designing the inhibitor and to understand the cellular mechanism of its cytotoxicity. Current assay methods to analyze the LF activity have been based on a synthetic peptide, consisting of 15-20 residues around being cleaved. However, there are accumulating reports that the region distal to cleavage site is required for the LF-mediated proteolysis of substrate. In this study, we demonstrate the catalytic properties of LF, using the full-length native substrate, MEK. We described the catalytic properties of LF focused on the effects of the pH alteration, which was encountered during the endocytosis of lethal toxin, and of the requirement for metal ions. We present the first evidence that additional metal ions are required for the LF catalyzed hydrolysis of native substrate, and that the pH alteration causes a significant change of catalytic properties of LF.
Kim
Joungmok
J
Department of Chemistry, Hanyang University, Seoul 133-791, Republic of Korea.
Chai
Young-Gyu
YG
Yoon
Moon-Young
MY
D000942:Antigens, Bacterial; D001051:Apoenzymes / Q000378:metabolism; D001408:Bacillus anthracis; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002384:Catalysis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000737:chemistry / Q000378:metabolism; D020929:Mitogen-Activated Protein Kinase Kinases / Q000737:chemistry; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015032:Zinc / Q000737:chemistry / Q000378:metabolism
D000942:Antigens, Bacterial; D001051:Apoenzymes; D001427:Bacterial Toxins; D009842:Oligopeptides; D011994:Recombinant Proteins; C030325:anthrax toxin; D020929:Mitogen-Activated Protein Kinase Kinases; D008274:Magnesium; D015032:Zinc; D002118:Calcium