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An increase in surface hydrophobicity mediates chaperone activity in N-chlorinated RidA

Varatnitskaya, Marharyta; Fasel, Julia; Müller, Alexandra; Lupilov, Natalie; Shi, Yunlong; Fuchs, Kristin; Krewing, Marco; Jung, Christoph 1; Jacob, Timo 2; Sitek, Barbara; Bandow, Julia E.; Carroll, Kate S.; Hofmann, Eckhard; Leichert, Lars I.
1 Institut für Wirtschaftsinformatik und Marketing (IISM), Karlsruher Institut für Technologie (KIT)
2 Institut für Mechanische Verfahrenstechnik und Mechanik (MVM), Karlsruher Institut für Technologie (KIT)

Abstract:

Under physiological conditions, Escherichia coli RidA is an enamine/imine deaminase, which promotes the release of ammonia from reactive enamine/imine intermediates. However, when modified by hypochlorous acid (HOCl), it turns into a potent chaperone-like holdase that can effectively protect E. coli's proteome during oxidative stress. However, it is unknown, which residues need to be chlorinated for activation. Here, we employ a combination of LC-MS/MS analysis, a chemo-proteomic approach, and a mutagenesis study to identify residues responsible for RidA's chaperone-like function. Through LC-MS/MS of digested RidAHOCl, we obtained direct evidence of the chlorination of one arginine residue. To overcome the instability of the N-chloramine modification, we established a chemoproteomic approach using 5-(dimethylamino) naphthalene-1-sulfinic acid (DANSO$_{2}$H) as a probe to label N-chlorinated lysines. Using this probe, we were able to detect the N-chlorination of six additional lysine residues. Moreover, using a mutagenesis study to genetically probe the role of single arginine and lysine residues, we found that the removal of arginines R105 and/or R128 led to a substantial reduction of RidA$_{HOCl's}$ chaperone activity. ... mehr


Verlagsausgabe §
DOI: 10.5445/IR/1000150892
Veröffentlicht am 22.09.2022
Originalveröffentlichung
DOI: 10.1016/j.redox.2022.102332
Scopus
Zitationen: 2
Web of Science
Zitationen: 2
Dimensions
Zitationen: 3
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Mechanische Verfahrenstechnik und Mechanik (MVM)
Institut für Wirtschaftsinformatik und Marketing (IISM)
Publikationstyp Zeitschriftenaufsatz
Publikationsmonat/-jahr 07.2022
Sprache Englisch
Identifikator ISSN: 2213-2317
KITopen-ID: 1000150892
Erschienen in Redox Biology
Verlag Elsevier
Band 53
Seiten Art.-Nr.: 102332
Schlagwörter N-Chlorination; Oxidation; Oxidative stress; E. coli; Chaperone
Nachgewiesen in Web of Science
Dimensions
Scopus
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