Effect of pH on stability of anthrax lethal factor: correlation between denaturation and activity.
Anthrax is caused by Gram positive bacterium Bacillus anthracis. Pathogenesis is result of production of three protein components, protective antigen (PA), lethal factor (LF), and edema factor (EF). PA in combination with LF (lethal toxin) is lethal to animals, while PA in combination with EF (edema toxin), causes edema. PA, LF, and EF are very thermolabile. Differential scanning calorimetry (DSC) was used to unravel the energetics of LF denaturation as a function of pH ranging from 7.8 to 5.5. Transition temperature (T(m)) of LF was found to be approximately equal to 42 degrees C and onset of denaturation occurs at approximately equal to 30 degrees C. The ratio of calorimetric to van't Hoff's enthalpy was nearly equal to unity at pH 7.0, indicative of presence of single structural domain in LF at pH 7.0, unlike PA which has been structurally observed to consist of 4 domains. It was found by cytotoxicity studies using J774A.1 macrophage like cells that LF was most stable at pH approximately 6.5. This paper reports for the first time the denaturation of LF at different pH values at 37 degrees C and tries to establish a correlation between denaturation and loss of LF activity at different pH values.
Gupta
P
P
Centre for Biotechnology, Jawaharlal Nehru University, New Delhi 110067, India.
Singh
S
S
Tiwari
A
A
Bhat
R
R
Bhatnagar
R
R
D000818:Animals; D000942:Antigens, Bacterial; D001427:Bacterial Toxins / Q000378:metabolism* / Q000494:pharmacology; D002152:Calorimetry, Differential Scanning; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011489:Protein Denaturation; D013696:Temperature; D013816:Thermodynamics
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; C030325:anthrax toxin