TY - JOUR
T1 - Role of arginine 180 and glutamic acid 177 of ricin toxin a chain in enzymatic inactivation of ribosomes
AU - Frankel, Arthur
AU - Welsh, Phil
AU - Richardson, Jane
AU - Robertus, Jon D.
N1 - Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1990
Y1 - 1990
N2 - The gene for ricin toxin A chain was modified by site-specific mutagenesis to change arginine 180 to alanine, glutamine, methionine, lysine, or histidine. Separately, glutamic acid 177 was changed to alanine and glutamic acid 208 was changed to aspartic acid. Both the wild-type and mutant proteins were expressed in Escherichia coli and, when soluble, purified and tested quantitatively for enzyme activity. A positive charge at position 180 was found necessary for solubility of the protein and for enzyme activity. Similarly, a negative charge with a proper geometry in the vicinity of position 177 was critical for ricin toxin A chain catalysis. When glutamic acid 177 was converted to alanine, nearby glutamic acid 208 could largely substitute for it. This observation provided valuable structural information concerning the nature of second-site mutations.
AB - The gene for ricin toxin A chain was modified by site-specific mutagenesis to change arginine 180 to alanine, glutamine, methionine, lysine, or histidine. Separately, glutamic acid 177 was changed to alanine and glutamic acid 208 was changed to aspartic acid. Both the wild-type and mutant proteins were expressed in Escherichia coli and, when soluble, purified and tested quantitatively for enzyme activity. A positive charge at position 180 was found necessary for solubility of the protein and for enzyme activity. Similarly, a negative charge with a proper geometry in the vicinity of position 177 was critical for ricin toxin A chain catalysis. When glutamic acid 177 was converted to alanine, nearby glutamic acid 208 could largely substitute for it. This observation provided valuable structural information concerning the nature of second-site mutations.
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U2 - 10.1128/MCB.10.12.6257
DO - 10.1128/MCB.10.12.6257
M3 - Article
C2 - 1978925
AN - SCOPUS:0025224121
SN - 0270-7306
VL - 10
SP - 6257
EP - 6263
JO - Molecular and cellular biology
JF - Molecular and cellular biology
IS - 12
ER -