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Title: Crystal structure of the Entamoeba histolytica RNA lariat debranching enzyme EhDbr1 reveals a catalytic Zn2+/Mn2+ heterobinucleation

Abstract

Here, the RNA lariat debranching enzyme, Dbr1, is a metallophosphoesterase that cleaves 2'-5' phosphodiester bonds within intronic lariats. Previous reports have indicated that Dbr1 enzymatic activity is supported by diverse metal ions including Ni2+, Mn2+, Mg2+, Fe2+, and Zn2+. While in initial structures of the Entamoeba histolytica Dbr1 only one of the two catalytic metal-binding sites were observed to be occupied (with a Mn2+ ion), recent structures determined a Zn2+/Fe2+ heterobinucleation. We solved a high-resolution X-ray crystal structure (1.8 Å) of the E. histolytica Dbr1 and determined a Zn2+/Mn2+ occupancy. ICP-AES corroborate this finding, and in vitro debranching assays with fluorescently labeled branched substrates confirm activity.

Authors:
 [1];  [2];  [2];  [2];  [3];  [4]
  1. Harvard Medical School, Boston, MA (United States)
  2. Carnegie Mellon Univ., Pittsburgh, PA (United States)
  3. Brookhaven National Lab. (BNL), Upton, NY (United States)
  4. Butler Univ., Indianapolis, IN (United States)
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1436261
Report Number(s):
BNL-203614-2018-JAAM
Journal ID: ISSN 0014-5793
Grant/Contract Number:  
SC0012704
Resource Type:
Accepted Manuscript
Journal Name:
FEBS Letters
Additional Journal Information:
Journal Volume: 591; Journal Issue: 13; Journal ID: ISSN 0014-5793
Publisher:
Federation of European Biochemical Societies
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE; heterobinucleation; ICP-AES; metallophosphoesterase

Citation Formats

Ransey, Elizabeth, Paredes, Eduardo, Dey, Sourav K., Das, Subha R., Heroux, Annie, and Macbeth, Mark R. Crystal structure of the Entamoeba histolytica RNA lariat debranching enzyme EhDbr1 reveals a catalytic Zn2+/Mn2+ heterobinucleation. United States: N. p., 2017. Web. doi:10.1002/1873-3468.12677.
Ransey, Elizabeth, Paredes, Eduardo, Dey, Sourav K., Das, Subha R., Heroux, Annie, & Macbeth, Mark R. Crystal structure of the Entamoeba histolytica RNA lariat debranching enzyme EhDbr1 reveals a catalytic Zn2+/Mn2+ heterobinucleation. United States. doi:10.1002/1873-3468.12677.
Ransey, Elizabeth, Paredes, Eduardo, Dey, Sourav K., Das, Subha R., Heroux, Annie, and Macbeth, Mark R. Wed . "Crystal structure of the Entamoeba histolytica RNA lariat debranching enzyme EhDbr1 reveals a catalytic Zn2+/Mn2+ heterobinucleation". United States. doi:10.1002/1873-3468.12677. https://www.osti.gov/servlets/purl/1436261.
@article{osti_1436261,
title = {Crystal structure of the Entamoeba histolytica RNA lariat debranching enzyme EhDbr1 reveals a catalytic Zn2+/Mn2+ heterobinucleation},
author = {Ransey, Elizabeth and Paredes, Eduardo and Dey, Sourav K. and Das, Subha R. and Heroux, Annie and Macbeth, Mark R.},
abstractNote = {Here, the RNA lariat debranching enzyme, Dbr1, is a metallophosphoesterase that cleaves 2'-5' phosphodiester bonds within intronic lariats. Previous reports have indicated that Dbr1 enzymatic activity is supported by diverse metal ions including Ni2+, Mn2+, Mg2+, Fe2+, and Zn2+. While in initial structures of the Entamoeba histolytica Dbr1 only one of the two catalytic metal-binding sites were observed to be occupied (with a Mn2+ ion), recent structures determined a Zn2+/Fe2+ heterobinucleation. We solved a high-resolution X-ray crystal structure (1.8 Å) of the E. histolytica Dbr1 and determined a Zn2+/Mn2+ occupancy. ICP-AES corroborate this finding, and in vitro debranching assays with fluorescently labeled branched substrates confirm activity.},
doi = {10.1002/1873-3468.12677},
journal = {FEBS Letters},
number = 13,
volume = 591,
place = {United States},
year = {2017},
month = {5}
}

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