Alkylation of Escherichia coli Thioredoxin by S-(2-Chloroethyl)glutathione and Identification of the Adduct on the Active Site Cysteine-32 by Mass Spectrometry

1995 ◽  
Vol 8 (7) ◽  
pp. 934-941 ◽  
Author(s):  
John C. L. Erve ◽  
Elisabeth Barofsky ◽  
Douglas F. Barofsky ◽  
Max L. Deinzer ◽  
Donald J. Reed
Keyword(s):  
Mass Spectrometry ◽  
Escherichia Coli ◽  
Active Site ◽  
Active Site Cysteine

scholarly journals Characterization of a 12-Kilodalton Rhodanese Encoded byglpE of Escherichia coli and Its Interaction with Thioredoxin

2000 ◽  
Vol 182 (8) ◽  
pp. 2277-2284 ◽  
Author(s):  
W. Keith Ray ◽  
Gang Zeng ◽  
M. Benjamin Potters ◽  
Aqil M. Mansuri ◽  
Timothy J. Larson
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Active Site ◽  
Amino Acid Sequences ◽  
E Coli ◽  
Active Site Cysteine ◽  
Clusters Of Orthologous Groups ◽  
Thioredoxin 1 ◽  
First Time

ABSTRACT Rhodaneses catalyze the transfer of the sulfane sulfur from thiosulfate or thiosulfonates to thiophilic acceptors such as cyanide and dithiols. In this work, we define for the first time the gene, and hence the amino acid sequence, of a 12-kDa rhodanese fromEscherichia coli. Well-characterized rhodaneses are comprised of two structurally similar ca. 15-kDa domains. Hence, it is thought that duplication of an ancestral rhodanese gene gave rise to the genes that encode the two-domain rhodaneses. The glpEgene, a member of the sn-glycerol 3-phosphate (glp) regulon of E. coli, encodes the 12-kDa rhodanese. As for other characterized rhodaneses, kinetic analysis revealed that catalysis by purified GlpE occurs by way of an enzyme-sulfur intermediate utilizing a double-displacement mechanism requiring an active-site cysteine. TheKm s for SSO3 2− and CN− were 78 and 17 mM, respectively. The apparent molecular mass of GlpE under nondenaturing conditions was 22.5 kDa, indicating that GlpE functions as a dimer. GlpE exhibited ak cat of 230 s−1. Thioredoxin 1 from E. coli, a small multifunctional dithiol protein, served as a sulfur acceptor substrate for GlpE with an apparentKm of 34 μM when thiosulfate was near itsKm , suggesting that thioredoxin 1 or related dithiol proteins could be physiological substrates for sulfurtransferases. The overall degree of amino acid sequence identity between GlpE and the active-site domain of mammalian rhodaneses is limited (∼17%). This work is significant because it begins to reveal the variation in amino acid sequences present in the sulfurtransferases. GlpE is the first among the 41 proteins in COG0607 (rhodanese-related sulfurtransferases) of the database Clusters of Orthologous Groups of proteins (http://www.ncbi.nlm.nih.gov/COG/ ) for which sulfurtransferase activity has been confirmed.

The C-Terminal Active Site Cysteine of Escherichia coli Glutaredoxin 1 Determines the Glutathione Specificity of the Second Step of Peptide Deglutathionylation

2009 ◽  
Vol 11 (8) ◽  
pp. 1819-1828 ◽  
Author(s):  
Mirva J. Saaranen ◽  
Kirsi E. H. Salo ◽  
Maria K. Latva-Ranta ◽  
Vuokko L. Kinnula ◽  
Lloyd W. Ruddock
Keyword(s):  
Escherichia Coli ◽  
Active Site ◽  
Second Step ◽  
Active Site Cysteine

scholarly journals Conformation, stability, and active-site cysteine titrations of Escherichia coli D26A thioredoxin probed by Raman spectroscopy

1998 ◽  
Vol 7 (1) ◽  
pp. 193-200 ◽  
Author(s):  
Stanislav Vohník ◽  
Roman Tuma ◽  
George J. Thomas ◽  
Chad Hanson ◽  
James A. Fuchs ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Raman Spectroscopy ◽  
Active Site ◽  
Active Site Cysteine ◽  
Conformation Stability

scholarly journals Identification of the l,d-Transpeptidases for Peptidoglycan Cross-Linking in Escherichia coli

2008 ◽  
Vol 190 (13) ◽  
pp. 4782-4785 ◽  
Author(s):  
Sophie Magnet ◽  
Lionel Dubost ◽  
Arul Marie ◽  
Michel Arthur ◽  
Laurent Gutmann
Keyword(s):  
Escherichia Coli ◽  
Active Site ◽  
Binding Proteins ◽  
Cross Linking ◽  
Peptide Bonds ◽  
Penicillin Binding Proteins ◽  
Active Site Cysteine ◽  
The Third ◽  
Family Form ◽  
Cross Links

ABSTRACT Three active-site cysteine l,d-transpeptidases can individually anchor the Braun lipoprotein to the Escherichia coli peptidoglycan. We show here that two additional enzymes of the same family form peptide bonds between the third residues of peptidoglycan stems, generating meso-DAP3→meso-DAP3 unusual cross-links. This activity partially replaces the d,d-transpeptidase activity of penicillin-binding proteins.

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