Evidence map›Paper›PMID 41820804›Full record

ArticleProtein science : a publication of the Protein Society2026

Accessing intractable, phosphorylated intrinsically disordered proteins via a protease-cleavable inclusion body tag.

Cat Hoang Vesely, Stanislau Stanisheuski, Edward Lien, Patrick Reardon, Peter J Chung, Ryan A Mehl, Richard B Cooley

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Protein science : a publication of the Protein Society, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Cat Hoang VeselyDepartment of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon, USA.
Stanislau StanisheuskiDepartment of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon, USA.
Edward LienDepartment of Physics, Department of Chemistry, Mann Department of Biomedical Engineering, University of Southern California, Los Angeles, California, USA.
Patrick ReardonDepartment of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon, USA.
Peter J ChungDepartment of Physics, Department of Chemistry, Mann Department of Biomedical Engineering, University of Southern California, Los Angeles, California, USA.
Ryan A MehlDepartment of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon, USA.
Richard B CooleyDepartment of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon, USA.ORCID https://orcid.org/0000-0003-3928-2757

Funding

The GCE4All Center: Unleashing the Potential of Genetic Code Expansion for Biomedical ResearchRM1GM144227 · NIGMS · OREGON STATE UNIVERSITY · PI RYAN A MEHL · 2022 to 2026
$6.2M
Equipment supplement: Defining Roles Of NitroTyrosine In Disease Via Genetic Code ExpansionR01GM114653 · NIGMS · OREGON STATE UNIVERSITY · PI MEHL, RYAN A · 2015 to 2024
$2.6M
The molecular determinants of surface-templated self-association of intrinsically disordered proteinsR35GM150716 · NIGMS · UNIVERSITY OF SOUTHERN CALIFORNIA · PI Peter J Chung · 2023 to 2026
$1.7M
Acquisition of a 700 MHz NMR SpectrometerS10OD018518 · OD · OREGON STATE UNIVERSITY · PI BARBAR, ELISAR J · 2015 to 2015
$1.3M
Epstein Breakthrough Alzheimer's Research FundM. J. Murdock Charitable Trust 2014162National Science Foundation 2320189NIGMS NIH HHS R01 GM114653NIGMS NIH HHS R35 GM150716NIGMS NIH HHS RM1 GM144227NIGMS NIH HHS RM1-GM144227NIH HHS 1S10OD018518NIH HHS 1S10OD02011NIH HHS 5R35GM150716NIH HHS S10 OD018518
6 · The paper itself

Abstract

Phosphorylated intrinsically disordered proteins (IDPs), such as Bcl2-associated agonist of cell death (BAD) and Tau, play critical roles in apoptosis and neurodegeneration, yet their instability during recombinant expression due to degradation, aggregation, and dephosphorylation, has limited structural and mechanistic studies. Here, we introduce the PINBody tag (PTM IDP Inclusion Body), a versatile inclusion body (IB)-targeting fusion that protects IDPs and their post-translational modifications during expression in Escherichia coli. When coupled with genetic code expansion (GCE), PINBody enables site-specific incorporation of phosphoserine (pS) while shielding this labile modification from dephosphorylation. In contrast to most IB-tag systems that rely on harsh chemical cleavage (e.g., acid, cyanogen bromide, heat) or slow, protease-based methods with limited refolding efficiency and IDP compatibility, PINBody supports rapid, complete protease-mediated tag removal under mild, refolding-compatible conditions. We demonstrate the generality of this system by producing milligram-scale quantities of phosphorylated mouse BAD (pS136), a protein that has evaded effective recombinant expression for over two decades, and 2N4R Tau phosphorylated at S404. To demonstrate PINbody utility, we generated homogeneous

Indexed as

Inclusion BodiesIntrinsically Disordered ProteinsRecombinant Fusion Proteinstau ProteinsAnimalsEscherichia coliMicePhosphorylationProtein Processing, Post-TranslationalIntrinsically Disordered ProteinsRecombinant Fusion Proteinstau Proteinsgenetic code expansioninclusion body tagintrinsically disordered proteinsNMRphosphorylationTau

Identifiers

PMID41820804
PMCPMC13140281

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.