Site-directed mutagenesis and deletion of the carboxyl terminus of Escherichia coli ribonucleotide reductase protein R2. Effects on catalytic activity and subunit interaction

Biochemistry ◽  
1992 ◽  
Vol 31 (20) ◽  
pp. 4801-4807 ◽  
Author(s):  
Isabel Climent ◽  
Britt Marie Sjoeberg ◽  
Charles Y. Huang
Keyword(s):  
Escherichia Coli ◽  
Catalytic Activity ◽  
Ribonucleotide Reductase ◽  
Site Directed Mutagenesis ◽  
Carboxyl Terminus ◽  
Directed Mutagenesis ◽  
Subunit Interaction

scholarly journals Riboflavin Synthase ofEscherichia coli

2001 ◽  
Vol 276 (15) ◽  
pp. 11524-11530 ◽  
Author(s):  
Boris Illarionov ◽  
Kristina Kemter ◽  
Sabine Eberhardt ◽  
Gerald Richter ◽  
Mark Cushman ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Catalytic Activity ◽  
Amino Acid ◽  
Binding Sites ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Sequence Motif ◽  
Amino Acid Residues ◽  
Terminal Sequence ◽  
Substrate Binding Sites

Conserved amino acid residues of riboflavin synthase fromEscherichia coliwere modified by site-directed mutagenesis. Replacement or deletion of phenylalanine 2 afforded catalytically inactive proteins. S41A and H102Q mutants had substantially reduced reaction velocities. Replacements of various other conserved polar residues had little impact on catalytic activity.19F NMR protein perturbation experiments using a fluorinated intermediate analog suggest that the N-terminal sequence motif MFTG is part of one of the substrate-binding sites of the protein.

Site directed mutagenesis of the class I ribonucleotide reductase from Escherichia coli

1991 ◽  
Vol 43 (2-3) ◽  
pp. 525
Author(s):  
Britt-Marie Sjöberg ◽  
Fredrick de Maré ◽  
Isabel Climent ◽  
Margareta Karlsson ◽  
Åke Larsson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ribonucleotide Reductase ◽  
Site Directed Mutagenesis ◽  
Class I ◽  
Directed Mutagenesis

scholarly journals Overexpression, site-directed mutagenesis, and mechanism of Escherichia coli acid phosphatase

1992 ◽  
Vol 267 (32) ◽  
pp. 22830-22836 ◽  
Author(s):  
K Ostanin ◽  
E.H. Harms ◽  
P.E. Stevis ◽  
R Kuciel ◽  
M.M. Zhou ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Acid Phosphatase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis

scholarly journals A genetically detoxified derivative of heat-labile Escherichia coli enterotoxin induces neutralizing antibodies against the A subunit.

1994 ◽  
Vol 180 (6) ◽  
pp. 2147-2153 ◽  
Author(s):  
M Pizza ◽  
M R Fontana ◽  
M M Giuliani ◽  
M Domenighini ◽  
C Magagnoli ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cholera Toxin ◽  
Neutralizing Antibodies ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
E Coli ◽  
Heat Labile ◽  
A Subunit ◽  
Adp Ribosyltransferase ◽  
Derivatives Of

Escherichia coli enterotoxin (LT) and the homologous cholera toxin (CT) are A-B toxins that cause travelers' diarrhea and cholera, respectively. So far, experimental live and killed vaccines against these diseases have been developed using only the nontoxic B portion of these toxins. The enzymatically active A subunit has not been used because it is responsible for the toxicity and it is reported to induce a negligible titer of toxin neutralizing antibodies. We used site-directed mutagenesis to inactivate the ADP-ribosyltransferase activity of the A subunit and obtained nontoxic derivatives of LT that elicited a good titer of neutralizing antibodies recognizing the A subunit. These LT mutants and equivalent mutants of CT may be used to improve live and killed vaccines against cholera and enterotoxinogenic E. coli.

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