Evidence mapPaperPMID 39762443Full record

ReviewNature protocols2025

Direct and quantitative analysis of tRNA acylation using intact tRNA liquid chromatography-mass spectrometry.

Riley Fricke, Isaac Knudson, Cameron Verdayne Swenson, Sarah Smaga, Alanna Schepartz

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In one paragraph

Review in Nature protocols, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

0numbers the graph read from it
0cells of the map it votes in
6citing 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

6 citing papers in PubMed.

  1. Article
  2. Co-Translational Incorporation ofJournal of the American Chemical Society · 2026
    Article
  3. Article
  4. Review
  5. Review
  6. 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

5 authors.

Riley FrickeDepartment of Chemistry, University of California, Berkeley, CA, USA.ORCID 0000-0001-9463-6314
Isaac KnudsonDepartment of Chemistry, University of California, Berkeley, CA, USA.
Cameron Verdayne SwensonDepartment of Chemistry, University of California, Berkeley, CA, USA.ORCID 0000-0002-9896-3539
Sarah SmagaDepartment of Chemistry, University of California, Berkeley, CA, USA.ORCID 0000-0003-3469-2642
Alanna SchepartzDepartment of Chemistry, University of California, Berkeley, CA, USA. schepartz@berkeley.edu.ORCID 0000-0003-2127-3932

Funding

National Science Foundation (NSF) CHE-2002182
6 · The paper itself

Abstract

Aminoacyl-tRNA synthetases (aaRSs) provide an essential functional link between an mRNA sequence and the protein it encodes. aaRS enzymes catalyze a two-step chemical reaction that acylates specific tRNAs with a cognate α-amino acid. In addition to their role in translation, acylated tRNAs contribute to non-ribosomal natural product biosynthesis and are implicated in multiple human diseases. In synthetic biology, the acylation of tRNAs with a non-canonical α-amino acid or, more recently, a non-α-amino acid monomer is a critical first step in the incorporation of these monomers into proteins, where they can be used for fundamental and applied science. These endeavors all demand an understanding of aaRS activity and specificity. Here, we describe a liquid chromatography-mass spectrometry assay that directly monitors aaRS activity by detecting the intact acyl-tRNA product. After a simple tRNA acylation reaction workup, acyl- and non-acyl-tRNA molecules are resolved by using ion-pairing reverse-phase chromatography, and their exact masses are determined by using high-resolution time-of-flight mass spectrometry. Our assay is fast and simple, quantifies reaction yields as low as 0.23% and can also be used on tRNAs acylated with flexizyme to detect products that are undetectable by using standard techniques. The protocol requires basic expertise in molecular biology, liquid chromatography-mass spectrometry and RNase-free techniques. This protocol takes ≥5 h to complete, depending on the number of samples.

Indexed as

Amino Acyl-tRNA SynthetasesMass SpectrometryRNA, TransferTransfer RNA AminoacylationAcylationChromatography, LiquidLiquid Chromatography-Mass SpectrometryAmino Acyl-tRNA SynthetasesRNA, Transfer

Identifiers

What Socratic holds

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Registered trials

None linked

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 Socratic graph.