Acid induced unfolding of anthrax protective antigen.
Acidic pH plays an important role in the membrane insertion of protective antigen (PA) of anthrax toxin leading to the translocation of the catalytic moieties. The structural transitions occurring in PA as a consequence of change in pH were investigated by fluorescence and circular dichroism measurements. Our studies revealed the presence of two intermediates on-pathway of acid induced unfolding; one at pH 2.0 and other at pH 4-5. Intrinsic fluorescence measurements of these intermediates showed a red shift in the wavelength of emission maximum with a concomitant decrease in fluorescence intensity, indicative of the exposure of tryptophan residues to the bulk solvent. Furthermore, no significant change was seen in the secondary structure of PA at a pH of 2.0, as indicated by far UV-CD spectra. The low pH intermediate of PA was characterized using the hydrophobic dye, 8-anilino-1-naphthalenesulfonate, and was found to have properties similar to those of a molten globule state.
Institute of Genomics and Integrative Biology, Delhi, India.
Pradeep K
PK
Gupta
Raj K
RK
Kurupati
Harish
H
Chandra
Reetika
R
Gaur
Vibha
V
Tandon
Yogendra
Y
Singh
Kapil
K
Maithal
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D000942:Antigens, Bacterial / Q000737:chemistry*; D001427:Bacterial Toxins / Q000737:chemistry*; D002942:Circular Dichroism; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D000817:Anilino Naphthalenesulfonates; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; C030325:anthrax toxin; C027132:1-anilino-8-naphthalenesulfonate