Evidence map›Paper›PMID 41676931›Full record

ArticleProtein science : a publication of the Protein Society2026

Optimized histidine-specific isotopic labeling in proteins overproduced in Escherichia coli.

Rosalie L Dohmen, Sarah M Teeman, Riddhi Patel, Elizabeth Schneider, Ethan Sparks, Amber J Dohmen, Janet Rogers, Margaret A Eastman, Steven D Hartson, Wouter D Hoff and 1 more

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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. Not yet cited in PubMed.

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0citing papers in PubMed
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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

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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

0 citing papers in PubMed.

No citing paper in PubMed yet.

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

11 authors.

Rosalie L DohmenDepartment of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.ORCID https://orcid.org/0009-0009-4314-5561
Sarah M TeemanDepartment of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
Riddhi PatelDepartment of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
Elizabeth SchneiderDepartment of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
Ethan SparksDepartment of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
Amber J DohmenDepartment of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
Janet RogersDepartment of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, Oklahoma, USA.
Margaret A EastmanDepartment of Chemistry, Oklahoma State University, Stillwater, Oklahoma, USA.
Steven D HartsonDepartment of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, Oklahoma, USA.
Wouter D HoffDepartment of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
Aihua XieDepartment of Physics, Oklahoma State University, Stillwater, Oklahoma, USA.

Funding

Advanced Infrared Biology of Protein Structure & DynamicsR15GM144898 · NIGMS · OKLAHOMA STATE UNIVERSITY STILLWATER · PI XIE, AIHUA · 2022 to 2024
$538k
National Science Foundation DBI 1338097NIGMS NIH HHS R15 GM144898NIH HHS R15GM144898-1Oklahoma State UniversityOSU Niblack Research Scholarship
6 · The paper itself

Abstract

The use of specific isotope labeling methods has significantly enhanced structural and kinetic research in protein science using a variety of biophysical techniques. Histidine plays key roles in a wide range of protein functions: enzymatic catalysis, metal ion coordination, protein stability, and redox reactions. In this article, we report a method for optimized histidine-specific isotopic labeling in proteins overproduced in Escherichia coli. We developed and validated this method using photoactive yellow protein (PYP) from Halorhodospira halophila, a prototype of the Per-Arnt-Sim (PAS) domain superfamily that contains only two native histidine residues. We tested both the efficiency of isotope editing of histidine and the degree of isotope scrambling after protein overproduction in E. coli grown in defined growth medium containing the 20 amino acids. Using varied amounts of isotope-edited histidine in the range of 0, 8, 16, 32, 64, and 128 mg/L of cell culture, we found that 32 mg of histidine per liter of cell culture is sufficient for over 98% of isotope labeling using double ionization mass spectroscopy measurements. Analysis of isotope envelopes of mass spectra of intact PYP are consistent with the absence of isotope scrambling, as are nuclear magnetic resonance spectra that show only resonances from the labeled histidine residues. In addition, we use Fourier transform infrared spectroscopy to demonstrate that these protein samples allow the detection of vibrational modes shifted upon histidine-specific isotope labeling. This method is widely applicable to histidine-specific isotope labeling of other proteins upon overexpression in E. coli.

Indexed as

Bacterial ProteinsEscherichia coliHalorhodospira halophilaHistidineIsotope LabelingPhotoreceptors, MicrobialRecombinant ProteinsBacterial ProteinsHistidinephotoactive yellow protein, BacteriaPhotoreceptors, MicrobialRecombinant ProteinsFourier transform infrared (FTIR) spectroscopyisotope editingLC–MS/MSnuclear magnetic resonance (NMR)PAS domain proteinsphotoactive yellow proteinresidue saturation

Identifiers

PMID41676931
PMCPMC12895364

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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.