Investigation of the role of the disulphide bond in the activity and structure of staphylococcal enterotoxin C1.
The goal of this study was to investigate the role of the disulphide bond of staphylococcal enterotoxin C1 (SEC1) in the structure and activity of the toxin. Mutants unable to form a disulphide bond were generated by substituting alanine or serine for cysteine at positions 93 and/or 110. Although we did not directly investigate the residues between the disulphide linkage, tryptic lability showed that significant native structure in the cystine loop is preserved in the absence of covalent bonding between residues 93 and 110. Since no correlation was observed between the behaviour of these mutants with regard to toxin stability, emesis and T cell proliferation we conclude that SEC1-induced emesis and T cell proliferation are dependent on separate regions of the molecule. The disulphide bond itself is not an absolute requirement for either activity. However, conformation within or adjacent to the loop is important for emesis. Although mutants with alanine substitutions were not emetic, those with serine substitutions retained this activity, suggesting that the disulphide linkage stabilizes a crucial conformation but can be replaced by residues which hydrogen bond.
Hovde
C J
CJ
Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow 83843.
Marr
J C
JC
Hoffmann
M L
ML
Hackett
S P
SP
Chi
Y I
YI
Crum
K K
KK
Stevens
D L
DL
Stauffacher
C V
CV
Bohach
G A
GA
D000595:Amino Acid Sequence; D000818:Animals; D003553:Cystine / Q000502:physiology*; D016207:Cytokines / Q000096:biosynthesis; D004768:Enterotoxins / Q000737:chemistry* / Q000494:pharmacology / Q000633:toxicity*; D006801:Humans; D006860:Hydrogen Bonding; D008213:Lymphocyte Activation / Q000187:drug effects*; D008254:Macaca nemestrina; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011817:Rabbits; D012772:Shock, Septic / Q000139:chemically induced / Q000209:etiology; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000187:drug effects; D014357:Trypsin / Q000378:metabolism; D014839:Vomiting / Q000139:chemically induced*
D016207:Cytokines; D004768:Enterotoxins; C034959:enterotoxin C, staphylococcal; D003553:Cystine; D014357:Trypsin