skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Biochemical and Structural Analysis of an Eis Family Aminoglycoside Acetyltransferase from Bacillus anthracis

Abstract

Proteins from the enhanced intracellular survival (Eis) family are versatile acetyltransferases that acetylate amines at multiple positions of several aminoglycosides (AGs). Their upregulation confers drug resistance. Homologues of Eis are present in diverse bacteria, including many pathogens. Eis from Mycobacterium tuberculosis (Eis_Mtb) has been well characterized. In this study, we explored the AG specificity and catalytic efficiency of the Eis family protein from Bacillus anthracis (Eis_Ban). Kinetic analysis of specificity and catalytic efficiency of acetylation of six AGs indicates that Eis_Ban displays significant differences from Eis_Mtb in both substrate binding and catalytic efficiency. The number of acetylated amines was also different for several AGs, indicating a distinct regiospecificity of Eis_Ban. Furthermore, most recently identified inhibitors of Eis_Mtb did not inhibit Eis_Ban, underscoring the differences between these two enzymes. To explain these differences, we determined an Eis_Ban crystal structure. The comparison of the crystal structures of Eis_Ban and Eis_Mtb demonstrates that critical residues lining their respective substrate binding pockets differ substantially, explaining their distinct specificities. Our results suggest that acetyltransferases of the Eis family evolved divergently to garner distinct specificities while conserving catalytic efficiency, possibly to counter distinct chemical challenges. The unique specificity features of these enzymes can be utilized asmore » tools for developing AGs with novel modifications and help guide specific AG treatments to avoid Eis-mediated resistance.« less

Authors:
; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Institutes of Health (NIH); USDOE Office of Science - Office of Biological and Environmental Research; University of Kentucky
OSTI Identifier:
1362300
DOE Contract Number:  
AC02-06CH11357
Resource Type:
Journal Article
Journal Name:
Biochemistry
Additional Journal Information:
Journal Volume: 54; Journal Issue: 20; Journal ID: ISSN 0006-2960
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English

Citation Formats

Green, Keith D., Biswas, Tapan, Chang, Changsoo, Wu, Ruiying, Chen, Wenjing, Janes, Brian K., Chalupska, Dominika, Gornicki, Piotr, Hanna, Philip C., Tsodikov, Oleg V., Joachimiak, Andrzej, and Garneau-Tsodikova, Sylvie. Biochemical and Structural Analysis of an Eis Family Aminoglycoside Acetyltransferase from Bacillus anthracis. United States: N. p., 2015. Web. doi:10.1021/acs.biochem.5b00244.
Green, Keith D., Biswas, Tapan, Chang, Changsoo, Wu, Ruiying, Chen, Wenjing, Janes, Brian K., Chalupska, Dominika, Gornicki, Piotr, Hanna, Philip C., Tsodikov, Oleg V., Joachimiak, Andrzej, & Garneau-Tsodikova, Sylvie. Biochemical and Structural Analysis of an Eis Family Aminoglycoside Acetyltransferase from Bacillus anthracis. United States. doi:10.1021/acs.biochem.5b00244.
Green, Keith D., Biswas, Tapan, Chang, Changsoo, Wu, Ruiying, Chen, Wenjing, Janes, Brian K., Chalupska, Dominika, Gornicki, Piotr, Hanna, Philip C., Tsodikov, Oleg V., Joachimiak, Andrzej, and Garneau-Tsodikova, Sylvie. Tue . "Biochemical and Structural Analysis of an Eis Family Aminoglycoside Acetyltransferase from Bacillus anthracis". United States. doi:10.1021/acs.biochem.5b00244.
@article{osti_1362300,
title = {Biochemical and Structural Analysis of an Eis Family Aminoglycoside Acetyltransferase from Bacillus anthracis},
author = {Green, Keith D. and Biswas, Tapan and Chang, Changsoo and Wu, Ruiying and Chen, Wenjing and Janes, Brian K. and Chalupska, Dominika and Gornicki, Piotr and Hanna, Philip C. and Tsodikov, Oleg V. and Joachimiak, Andrzej and Garneau-Tsodikova, Sylvie},
abstractNote = {Proteins from the enhanced intracellular survival (Eis) family are versatile acetyltransferases that acetylate amines at multiple positions of several aminoglycosides (AGs). Their upregulation confers drug resistance. Homologues of Eis are present in diverse bacteria, including many pathogens. Eis from Mycobacterium tuberculosis (Eis_Mtb) has been well characterized. In this study, we explored the AG specificity and catalytic efficiency of the Eis family protein from Bacillus anthracis (Eis_Ban). Kinetic analysis of specificity and catalytic efficiency of acetylation of six AGs indicates that Eis_Ban displays significant differences from Eis_Mtb in both substrate binding and catalytic efficiency. The number of acetylated amines was also different for several AGs, indicating a distinct regiospecificity of Eis_Ban. Furthermore, most recently identified inhibitors of Eis_Mtb did not inhibit Eis_Ban, underscoring the differences between these two enzymes. To explain these differences, we determined an Eis_Ban crystal structure. The comparison of the crystal structures of Eis_Ban and Eis_Mtb demonstrates that critical residues lining their respective substrate binding pockets differ substantially, explaining their distinct specificities. Our results suggest that acetyltransferases of the Eis family evolved divergently to garner distinct specificities while conserving catalytic efficiency, possibly to counter distinct chemical challenges. The unique specificity features of these enzymes can be utilized as tools for developing AGs with novel modifications and help guide specific AG treatments to avoid Eis-mediated resistance.},
doi = {10.1021/acs.biochem.5b00244},
journal = {Biochemistry},
issn = {0006-2960},
number = 20,
volume = 54,
place = {United States},
year = {2015},
month = {5}
}