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PMID: 2689871 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Role of glutamic acid 177 of the ricin toxin A chain in enzymatic inactivation of ribosomes.

Molecular and cellular biology ·Vol. 9 ·No. 11 ·1989-11-00 ·Pages 5012-21

Schlossman D, Withers D, Welsh P, Alexander A, Robertus J, Frankel A

Abstract

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.

MeSH Terms
Blotting, Western Cloning, Molecular Electrophoresis, Polyacrylamide Gel Escherichia coli/genetics Gene Expression Glutamates/metabolism Immunoenzyme Techniques Macromolecular Substances Mutation Ribosomes/metabolism Ricin/genetics,isolation & purification,metabolism Yeasts/genetics
Chemicals
Glutamates Macromolecular Substances Ricin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Schlossman D
Department of Medicine, Duke University Medical Center, Durham, North Carolina 22710.
Withers D
Welsh P
Alexander A
Robertus J
Frankel A
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29 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.9.11.5012-5021.1989
Published
1989-11-00
Pages
5012-21
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC363653
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.9.11.5012-5021.1989 PMC Full Text View journal details
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