Preprint
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- Not peer reviewed
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(Preview, Version of record, pdf, 592.7KB, Terms of use)
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- Preprint server copy:
- 10.1101/2023.02.27.529914
Authors
+ European Research Council
More from this funder
- Funder identifier:
- https://ror.org/0472cxd90
- Funding agency for:
- van der Hoorn, RAL
- Grant:
- 101019324
- Programme:
- ExtraImmune
+ Biotechnology and Biological Sciences Research Council
More from this funder
- Funder identifier:
- https://ror.org/00cwqg982
- Funding agency for:
- Sanguankiattichai, N
- Grant:
- BB/T015128/1
- Programme:
- Galactosyrin
+ Science Achievement Scholarship of Thailand
More from this funder
- Funding agency for:
- Longsaward, R
- Preprint server:
- bioRxiv
- Publication date:
- 2023-02-28
- DOI:
- Language:
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English
- Keywords:
- Pubs id:
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1331446
- Local pid:
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pubs:1331446
- Deposit date:
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2025-04-29
Terms of use
- Copyright holder:
- Longsaward et al.
- Copyright date:
- 2023
- Rights statement:
- The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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