Identification of amino acid residues of anthrax protective antigen involved in binding with lethal factor.
Protective antigen (PA) and lethal factor (LF) are the two components of anthrax lethal toxin. PA is responsible for the translocation of LF to the cytosol. The binding of LF to cell surface receptor-bound PA is a prerequisite for the formation of lethal toxin. It has been hypothesized that hydrophobic residues P184, L187, F202, L203, P205, I207, I210, W226, and F236 of domain 1b of PA play an important role in the binding of PA to LF. These residues are normally buried in the 83-kDA version of PA, PA83, as determined by the crystal structure of PA. However, they become exposed due to the conformational change brought about by the cleavage of PA83 to PA63 by a cell surface protease. Mutation of the above-mentioned residues to alanine resulted in mutant proteins that were able to bind to the cell surface receptors and also to be specifically cleaved by the cellular proteases. All the mutant proteins except the F202A, L203A, P205A, and I207A mutants were able to bind to LF and were also toxic to macrophage cells in combination with LF. It was concluded that residues 202, 203, 205, and 207 of PA are essential for the binding of LF to PA.
Chauhan
Vibha
V
Centre For Biotechnology, Jawaharlal Nehru University, New Delhi 110067, India.
Bhatnagar
Rakesh
R
D020816:Amino Acid Motifs; D000818:Animals; D000942:Antigens, Bacterial; D001408:Bacillus anthracis / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001665:Binding Sites; D002460:Cell Line; D003603:Cytotoxins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli; D015870:Gene Expression; D051379:Mice; D016297:Mutagenesis, Site-Directed; D011956:Receptors, Cell Surface / Q000378:metabolism; D012996:Solutions; D014357:Trypsin / Q000378:metabolism
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D003603:Cytotoxins; D011956:Receptors, Cell Surface; D012996:Solutions; C030325:anthrax toxin; D014357:Trypsin