A multidomain PARP14 construct suitable for bacterial expression
(2024) In Protein Expression and Purification 224.- Abstract
Poly-ADP-ribose polymerase-14 (PARP14) can modify proteins and nucleic acids by the reversible addition of a single ADP-ribose molecule. Aberrant PARP14 functions have been related to cancer and inflammation, and its domains are involved in processes related to viral infection. Previous research indicates that PARP14 functions might be mediated via a multitude of target proteins. In vitro studies of this large multidomain enzyme have been complicated by difficulties to obtain biochemical quantities of pure protein. Here we present a strategy that allows bacterial expression and purification of a functional multidomain construct of PARP14. We substituted an internal KH domain and its neighboring unstructured region with a SUMO domain to... (More)
Poly-ADP-ribose polymerase-14 (PARP14) can modify proteins and nucleic acids by the reversible addition of a single ADP-ribose molecule. Aberrant PARP14 functions have been related to cancer and inflammation, and its domains are involved in processes related to viral infection. Previous research indicates that PARP14 functions might be mediated via a multitude of target proteins. In vitro studies of this large multidomain enzyme have been complicated by difficulties to obtain biochemical quantities of pure protein. Here we present a strategy that allows bacterial expression and purification of a functional multidomain construct of PARP14. We substituted an internal KH domain and its neighboring unstructured region with a SUMO domain to obtain a protein construct that encompasses three macrodomains, a WWE domain, and a PARP catalytic domain. We show that the resulting construct retains both ADP-ribosyltransferase and de-MARylase activities. This construct will be useful in structural and functional studies of PARP14.
(Less)
- author
- Chatzicharalampous, Constantinos
LU
and Schüler, Herwig LU
- organization
- publishing date
- 2024-08-19
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Protein Expression and Purification
- volume
- 224
- article number
- 106580
- pages
- 8 pages
- publisher
- Academic Press
- external identifiers
-
- scopus:85201504050
- pmid:39154924
- ISSN
- 1046-5928
- DOI
- 10.1016/j.pep.2024.106580
- language
- English
- LU publication?
- yes
- additional info
- Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.
- id
- d3ec38f3-ff40-4eb0-9b9a-bff8b7051759
- date added to LUP
- 2024-08-24 00:14:56
- date last changed
- 2024-09-07 05:28:01
@article{d3ec38f3-ff40-4eb0-9b9a-bff8b7051759, abstract = {{<p>Poly-ADP-ribose polymerase-14 (PARP14) can modify proteins and nucleic acids by the reversible addition of a single ADP-ribose molecule. Aberrant PARP14 functions have been related to cancer and inflammation, and its domains are involved in processes related to viral infection. Previous research indicates that PARP14 functions might be mediated via a multitude of target proteins. In vitro studies of this large multidomain enzyme have been complicated by difficulties to obtain biochemical quantities of pure protein. Here we present a strategy that allows bacterial expression and purification of a functional multidomain construct of PARP14. We substituted an internal KH domain and its neighboring unstructured region with a SUMO domain to obtain a protein construct that encompasses three macrodomains, a WWE domain, and a PARP catalytic domain. We show that the resulting construct retains both ADP-ribosyltransferase and de-MARylase activities. This construct will be useful in structural and functional studies of PARP14.</p>}}, author = {{Chatzicharalampous, Constantinos and Schüler, Herwig}}, issn = {{1046-5928}}, language = {{eng}}, month = {{08}}, publisher = {{Academic Press}}, series = {{Protein Expression and Purification}}, title = {{A multidomain PARP14 construct suitable for bacterial expression}}, url = {{http://dx.doi.org/10.1016/j.pep.2024.106580}}, doi = {{10.1016/j.pep.2024.106580}}, volume = {{224}}, year = {{2024}}, }