Hydrophobic residues Phe552, Phe554, Ile562, Leu566, and Ile574 are required for oligomerization of anthrax protective antigen.
Anthrax protective antigen (PA) plays a central role in facilitating the entry of active toxin components, namely, lethal factor and edema factor, into the cells. PA is also the main immunogen of both human and veterinary vaccine against anthrax. During host cell intoxication, protective antigen binds to the receptors on cell surface, gets proteolytically activated, oligomerizes to form a heptamer and binds to lethal factor or edema factor. The complex, formed by binding of lethal factor or edema factor to oligomerized PA, is internalized by receptor-mediated endocytosis. Acidification of the endosome results in the insertion of the heptamer into the membrane, thereby forming a pore through which lethal factor or edema factor can translocate into the cytosol. In this study we have identified hydrophobic residues, Phe552, Phe554, Ile562, Leu566, and Ile574, which are required for oligomerization of anthrax protective antigen. Mutation of these conserved residues to alanine impaired the oligomerization of protective antigen. Consequently, these mutants became nontoxic in combination with lethal factor and edema factor. Therapeutic importance of these mutants and their potential as vaccine candidates is discussed.
Ahuja
N
N
Centre for Biotechnology, Jawaharlal Nehru University, New Delhi 110067, India.
Kumar
P
P
Bhatnagar
R
R
D000409:Alanine / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000942:Antigens, Bacterial; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D001667:Binding, Competitive; D001704:Biopolymers / Q000378:metabolism / Q000494:pharmacology; D016466:CHO Cells / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D007532:Isoleucine / Q000235:genetics / Q000378:metabolism; D007930:Leucine / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010447:Peptide Hydrolases / Q000378:metabolism; D010649:Phenylalanine / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D001704:Biopolymers; D011956:Receptors, Cell Surface; C030325:anthrax toxin; D007532:Isoleucine; D010649:Phenylalanine; D010447:Peptide Hydrolases; D007930:Leucine; D000409:Alanine