title: Role of cysteine 41 of the A subunit of pertussis toxin.
abstract: The 2 cysteine residues present in the A subunit of pertussis toxin form a disulfide bond in the conformation of the toxin secreted from the bacteria. Previous studies have shown that reduction of this bond is necessary for activation of the enzyme. We have found that reduction of this bond also alters the conformation of the A subunit such that it no longer readily associates with the B oligomer of the toxin, a finding which may have implications concerning the form of the toxin found within the eukaryotic cell. In addition, we have demonstrated that reduction of the disulfide bond of the purified A subunit followed by treatment with sulfhydryl-modifying reagents such as N-ethylmaleimide or 5,5'-dithiobis-(2-nitrobenzoic acid) results in inhibition of the NAD glycohydrolase activity of the protein. When a tryptic fragment of the A subunit which contains only 1 of the cysteine residues (Cys-41) of the native protein was reacted with N-ethylmaleimide, the NAD glycohydrolase activity of this fragment was substantially reduced. These data indicate that Cys-41 may be in a region of the molecule which is critical for the enzymatic activity of the toxin.
authors.affiliation: Center for Biologies Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892.
authors.forename: D L
authors.initials: DL
authors.lastname: Burns
authors.forename: C R
authors.initials: CR
authors.lastname: Manclark
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003545:Cysteine; D004220:Disulfides / Q000378:metabolism; D005033:Ethylmaleimide / Q000494:pharmacology; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000378:metabolism; D008970:Molecular Weight; D009244:NAD+ Nucleosidase / Q000378:metabolism; D037342:Pertussis Toxin; D011487:Protein Conformation; D010566:Virulence Factors, Bordetella / Q000378:metabolism
chemical_list: D004220:Disulfides; D046911:Macromolecular Substances; D008565:Membrane Proteins; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin; D009244:NAD+ Nucleosidase; D003545:Cysteine; D005033:Ethylmaleimide