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Cautionary note on ribonuclease activity of recombinant PR-10 proteins

Abstract:
We studied the biochemical properties of three splicing isoforms of PR-10 from rubber tree (Hevea brasiliensis) and found that purified recombinant HbPR10 can cause RNA degradation in vitro, a well-known activity described for many PR-10 proteins. This ribonuclease activity was observed for all three HbPR10 splicing isoforms and is abolished by boiling. However, inclusion of a negative control proteins revealed that ribonuclease activity rather originates from RNases that are copurified from E. coli, which are overlooked by traditionally used controls such as heat inactivation, RNase inhibitors and negative control proteins obtained with different procedures. The crucial control proteins are missing for at least nine reports on ribonuclease activity in PR-10 proteins published by different laboratories worldwide, indicating that proper controls are frequently overlooked in ribonuclease assays. The raised cautionary note applies to several PR-10 proteins with proclaimed ribonuclease activities and call for the use of different assays and mutant PR-10 proteins as control.
Publication status:
Published
Peer review status:
Not peer reviewed

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Preprint server copy:
10.1101/2023.02.27.529914

Authors


More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Biology
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Biology
Role:
Author
ORCID:
0000-0003-4408-8262
More by this author
Role:
Author
ORCID:
0000-0001-7707-3687
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Biology
Oxford college:
Somerville College
Role:
Author
ORCID:
0000-0002-3692-7487


More from this funder
Funder identifier:
https://ror.org/0472cxd90
Funding agency for:
van der Hoorn, RAL
Grant:
101019324
Programme:
ExtraImmune
More from this funder
Funder identifier:
https://ror.org/00cwqg982
Funding agency for:
Sanguankiattichai, N
Grant:
BB/T015128/1
Programme:
Galactosyrin


Preprint server:
bioRxiv
Publication date:
2023-02-28
DOI:


Language:
English
Keywords:
Pubs id:
1331446
Local pid:
pubs:1331446
Deposit date:
2025-04-29

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