Evidence map›Paper›PMID 40650557›Full record

ArticleCytoskeleton (Hoboken, N.J.)2025

Purification, Fluorescent Labeling, and Detyrosination of Mammalian Cell Tubulin for Biochemical Assays.

Ezekiel C Thomas, Yang Yue, Morgan L Pimm, Takashi Hotta, Ryoma Ohi, Kristen J Verhey

Abstract read
In one paragraph

Article in Cytoskeleton (Hoboken, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing 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

5 citing papers in PubMed.

  1. Article
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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

6 authors.

Ezekiel C ThomasDepartment of Cell and Developmental Biology, University of Michigan, Ann Arbor, Michigan, USA.ORCID https://orcid.org/0000-0002-9810-0477
Yang YueDepartment of Cell and Developmental Biology, University of Michigan, Ann Arbor, Michigan, USA.ORCID https://orcid.org/0000-0002-8302-9778
Morgan L PimmDepartment of Cell and Developmental Biology, University of Michigan, Ann Arbor, Michigan, USA.ORCID https://orcid.org/0000-0001-6370-1435
Takashi HottaDepartment of Cell and Developmental Biology, University of Michigan, Ann Arbor, Michigan, USA.
Ryoma OhiDepartment of Cell and Developmental Biology, University of Michigan, Ann Arbor, Michigan, USA.
Kristen J VerheyDepartment of Cell and Developmental Biology, University of Michigan, Ann Arbor, Michigan, USA.ORCID https://orcid.org/0000-0001-9329-4981

Funding

Kinesin Motors and Microtubule-based TraffickingR35GM131744 · NIGMS · UNIVERSITY OF MICHIGAN AT ANN ARBOR · PI Kristen J. Verhey · 2019 to 2026
$6.3M
Centriole assembly and function for centrosome and cilia biologyR35GM140813 · NIGMS · UNIVERSITY OF COLORADO DENVER · PI CHAD G PEARSON · 2021 to 2026
$3.5M
Biology and Biochemistry of the Microtubule CytoskeletonR35GM153209 · NIGMS · UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR · PI Ryoma Ohi · 2024 to 2026
$1.0M
Development of live-cell probes to investigate tubulin post-translational modifications in neuronal regenerationR21NS132003 · NINDS · UNIVERSITY OF MICHIGAN AT ANN ARBOR · PI GIGER, ROMAN JENO, OHI, RYOMA · 2023 to 2024
$429k
Feedback mechanisms link tubulin PTMs to Golgi organizationF32GM157897 · NIGMS · UNIVERSITY OF MICHIGAN AT ANN ARBOR · PI Morgan Lynn Pimm · 2025 to 2026
$152k
American Cancer Society PF-24-1320851-01-CCBNIGMS NIH HHS F32 GM157897NIGMS NIH HHS R35 GM131744NIGMS NIH HHS R35 GM140813NIGMS NIH HHS R35 GM153209NIH HHS F32GM157897NIH HHS R35GM131744NIH HHS R35GM153209NINDS NIH HHS R21 NS132003
6 · The paper itself

Abstract

Microtubules play essential roles in numerous cellular processes. All microtubules are built from the protein tubulin, yet individual microtubules can differ spatially and temporally due to their tubulin isotype composition and post-translational modifications (PTMs). The tubulin code hypothesis posits that these differences can regulate microtubule function. However, investigating the properties of specific tubulin PTMs in vitro has been challenging because most reconstitution assays rely on tubulin purified from brain tissue that contains highly heterogeneous and modified microtubules. In this study, we present an optimized method for the purification of milligram quantities of unmodified tubulin from large-scale cultures of HeLa S3 cells. We also describe steps for efficient chemical labeling of tubulin and the generation of controlled tubulin PTMs. These tubulins can be used in microscopy or biochemistry-based experiments to investigate how the tubulin code influences microtubule properties and functions. Overall, our method is easily adaptable, highly reproducible, and broadly accessible to labs with general equipment.

Indexed as

HeLa tubulinmammalian cellsmicrotubulesprotein labelingsuspension cultureTOG affinity chromatographytubulin codetubulin purification

Identifiers

PMID40650557
PMCPMC12276855

What OpenQuestion holds

Textmetadata
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Read underepoch 390

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 OpenQuestion graph.