A biotin-HaloTag ligand enables efficient affinity capture of protein variants from live cells

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F25%3A00637714" target="_blank" >RIV/68081707:_____/25:00637714 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:14310/25:00142954 RIV/00159816:_____/25:00082440

Výsledek na webu

<a href="https://rupress.org/jcb/article-abstract/224/8/e202410025/278122/A-biotin-HaloTag-ligand-enables-efficient-affinity?redirectedFrom=fulltext" target="_blank" >https://rupress.org/jcb/article-abstract/224/8/e202410025/278122/A-biotin-HaloTag-ligand-enables-efficient-affinity?redirectedFrom=fulltext</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1083/jcb.202410025" target="_blank" >10.1083/jcb.202410025</a>

Jazyk výsledku

angličtina

Název v původním jazyce

A biotin-HaloTag ligand enables efficient affinity capture of protein variants from live cells

Popis výsledku v původním jazyce

HaloTag technology represents a versatile tool for studying proteins. Fluorescent HaloTag ligands employed in sequential labeling led to the discovery of distinct protein variants for histones, cohesins, and MCM complexes. However, an efficient biochemical approach to separate these distinct protein variants to study their biological functions is missing. Principally, being a gap in technology, the HaloTag toolbox lacks affinity ligands displaying good cell permeability and efficient affinity capture. Here, we describe the design, synthesis, and validation of a new cell-permeable biotin-HaloTag ligand, which allows rapid labeling of Halo-tagged proteins in live cells and their efficient separation using streptavidin pull-down. We provide a proof-of-concept application of how to use the herein-developed affinity ligand in sequential labeling to biochemically separate protein variants and study their biological properties. This approach enables to address fundamental questions concerning essential cellular processes, including genome duplication and chromatin maintenance.

Název v anglickém jazyce

A biotin-HaloTag ligand enables efficient affinity capture of protein variants from live cells

Popis výsledku anglicky

HaloTag technology represents a versatile tool for studying proteins. Fluorescent HaloTag ligands employed in sequential labeling led to the discovery of distinct protein variants for histones, cohesins, and MCM complexes. However, an efficient biochemical approach to separate these distinct protein variants to study their biological functions is missing. Principally, being a gap in technology, the HaloTag toolbox lacks affinity ligands displaying good cell permeability and efficient affinity capture. Here, we describe the design, synthesis, and validation of a new cell-permeable biotin-HaloTag ligand, which allows rapid labeling of Halo-tagged proteins in live cells and their efficient separation using streptavidin pull-down. We provide a proof-of-concept application of how to use the herein-developed affinity ligand in sequential labeling to biochemically separate protein variants and study their biological properties. This approach enables to address fundamental questions concerning essential cellular processes, including genome duplication and chromatin maintenance.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10601 - Cell biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Cell Biology

ISSN

0021-9525

e-ISSN

1540-8140

Svazek periodika

224

Číslo periodika v rámci svazku

8

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

26

Strana od-do

e202410025

Kód UT WoS článku

001530070500001

EID výsledku v databázi Scopus

2-s2.0-105011883583