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Title: Structural basis for the diversity of the mechanism of nucleotide hydrolysis by the aminoglycoside-2"-phosphotransferases

Abstract

Aminoglycoside phosphotransferases (APHs) are one of three families of aminoglycoside-modifying enzymes that confer high-level resistance to the aminoglycoside antibiotics via enzymatic modification. This has now rendered many clinically important drugs almost obsolete. The APHs specifically phosphorylate hydroxyl groups on the aminoglycosides using a nucleotide triphosphate as the phosphate donor. The APH(2′′) family comprises four distinct members, isolated primarily from Enterococcus sp., which vary in their substrate specificities and also in their preference for the phosphate donor (ATP or GTP). In this work, the structure of the ternary complex of APH(2′′)-IIIa with GDP and kanamycin was solved at 1.34 Å resolution and was compared with substrate-bound structures of APH(2′′)-Ia, APH(2′′)-IIa and APH(2′′)-IVa. In contrast to the case for APH(2′′)-Ia, where it was proposed that the enzyme-mediated hydrolysis of GTP is regulated by conformational changes in its N-terminal domain upon GTP binding, APH(2′′)-IIa, APH(2′′)-IIIa and APH(2′′)-IVa show no such regulatory mechanism, primarily owing to structural differences in the N-terminal domains of these enzymes.

Authors:
 [1];  [2];  [2];  [3];  [2]
  1. SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL); Stanford Univ., CA (United States)
  2. Univ. of Notre Dame, IN (United States). Dept. of Chemistry and Biochemistry
  3. SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Workforce Development for Teachers and Scientists (WDTS) (SC-27); National Institutes of Health (NIH)
OSTI Identifier:
1595141
Grant/Contract Number:  
[AC02-76SF00515; 2R01AI057393; P41GM103393]
Resource Type:
Accepted Manuscript
Journal Name:
Acta Crystallographica. Section D. Structural Biology
Additional Journal Information:
[ Journal Volume: 75; Journal Issue: 12]; Journal ID: ISSN 2059-7983
Publisher:
IUCr
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; crystal structure; aminoglycosides; phosphotransferases; antibiotic resistance

Citation Formats

Smith, Clyde A., Toth, Marta, Stewart, Nichole K., Maltz, Lauren, and Vakulenko, Sergei B. Structural basis for the diversity of the mechanism of nucleotide hydrolysis by the aminoglycoside-2"-phosphotransferases. United States: N. p., 2019. Web. doi:10.1107/S2059798319015079.
Smith, Clyde A., Toth, Marta, Stewart, Nichole K., Maltz, Lauren, & Vakulenko, Sergei B. Structural basis for the diversity of the mechanism of nucleotide hydrolysis by the aminoglycoside-2"-phosphotransferases. United States. doi:10.1107/S2059798319015079.
Smith, Clyde A., Toth, Marta, Stewart, Nichole K., Maltz, Lauren, and Vakulenko, Sergei B. Fri . "Structural basis for the diversity of the mechanism of nucleotide hydrolysis by the aminoglycoside-2"-phosphotransferases". United States. doi:10.1107/S2059798319015079.
@article{osti_1595141,
title = {Structural basis for the diversity of the mechanism of nucleotide hydrolysis by the aminoglycoside-2"-phosphotransferases},
author = {Smith, Clyde A. and Toth, Marta and Stewart, Nichole K. and Maltz, Lauren and Vakulenko, Sergei B.},
abstractNote = {Aminoglycoside phosphotransferases (APHs) are one of three families of aminoglycoside-modifying enzymes that confer high-level resistance to the aminoglycoside antibiotics via enzymatic modification. This has now rendered many clinically important drugs almost obsolete. The APHs specifically phosphorylate hydroxyl groups on the aminoglycosides using a nucleotide triphosphate as the phosphate donor. The APH(2′′) family comprises four distinct members, isolated primarily from Enterococcus sp., which vary in their substrate specificities and also in their preference for the phosphate donor (ATP or GTP). In this work, the structure of the ternary complex of APH(2′′)-IIIa with GDP and kanamycin was solved at 1.34 Å resolution and was compared with substrate-bound structures of APH(2′′)-Ia, APH(2′′)-IIa and APH(2′′)-IVa. In contrast to the case for APH(2′′)-Ia, where it was proposed that the enzyme-mediated hydrolysis of GTP is regulated by conformational changes in its N-terminal domain upon GTP binding, APH(2′′)-IIa, APH(2′′)-IIIa and APH(2′′)-IVa show no such regulatory mechanism, primarily owing to structural differences in the N-terminal domains of these enzymes.},
doi = {10.1107/S2059798319015079},
journal = {Acta Crystallographica. Section D. Structural Biology},
number = [12],
volume = [75],
place = {United States},
year = {2019},
month = {11}
}

Journal Article:
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