Role of glutamic acid 177 of the ricin toxin A chain in enzymatic inactivation of ribosomes.
The gene for the A chain of ricin toxin was fused to a beta-galactosidase marker cistron via a DNA sequence encoding a short collagen linker, and the tripartite fusion protein was expressed in Escherichia coli. Site-specific mutagenesis was used to change glutamic acid residue 177 to aspartic acid or alanine. When the mutant proteins were expressed, purified, and tested quantitatively for enzymatic activity, the carboxylate function at position 177 was found not to be absolutely essential for ricin toxin A-chain catalysis.
Schlossman
D
D
Department of Medicine, Duke University Medical Center, Durham, North Carolina 22710.
Withers
D
D
Welsh
P
P
Alexander
A
A
Robertus
J
J
Frankel
A
A
D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005971:Glutamates / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D046911:Macromolecular Substances; D009154:Mutation; D012270:Ribosomes / Q000378:metabolism*; D012276:Ricin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015003:Yeasts / Q000235:genetics
D005971:Glutamates; D046911:Macromolecular Substances; D012276:Ricin