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

Journal Article · · Acta Crystallographica. Section D. Structural Biology
 [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)
+ Show Author Affiliations

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.

Research Organization:
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Science (SC), Workforce Development for Teachers and Scientists (WDTS); National Institutes of Health (NIH)
Grant/Contract Number:
AC02-76SF00515; 2R01AI057393; P41GM103393
OSTI ID:
1595141
Journal Information:
Acta Crystallographica. Section D. Structural Biology, Vol. 75, Issue 12; ISSN 2059-7983
Publisher:
IUCrCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 1 work
Citation information provided by
Web of Science

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  • Byrnes, Laura J.; Badarau, Adriana; Vakulenko, Sergei B.
  • Acta Crystallographica Section F Structural Biology and Crystallization Communications, Vol. 64, Issue 2 https://doi.org/10.1107/S1744309108001450
journal January 2008
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59 BASIC BIOLOGICAL SCIENCES
crystal structure
aminoglycosides
phosphotransferases
antibiotic resistance