Fast Fluoroalkylation of Proteins Uncovers the Structure and Dynamics of Biological Macromolecules

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F21%3A00551051" target="_blank" >RIV/61388971:_____/21:00551051 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/86652036:_____/21:00556422 RIV/61388963:_____/21:00551051 RIV/00216208:11310/21:10437999

Výsledek na webu

<a href="https://pubs.acs.org/doi/10.1021/jacs.1c07771" target="_blank" >https://pubs.acs.org/doi/10.1021/jacs.1c07771</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/jacs.1c07771" target="_blank" >10.1021/jacs.1c07771</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Fast Fluoroalkylation of Proteins Uncovers the Structure and Dynamics of Biological Macromolecules

Popis výsledku v původním jazyce

Covalent labeling of proteins in combination with mass spectrometry has been established as a complementary technique to classical structural methods, such as X-ray, NMR, or cryogenic electron microscopy (Cryo-EM), used for protein structure determination. Although the current covalent labeling techniques enable the protein solvent accessible areas with sufficient spatial resolution to be monitored, there is still high demand for alternative, less complicated, and inexpensive approaches. Here, we introduce a new covalent labeling method based on fast fluoroalkylation of proteins (FFAP). FFAP uses fluoroalkyl radicals formed by reductive decomposition of Togni reagents with ascorbic acid to label proteins on a time scale of seconds. The feasibility of FFAP to effectively label proteins was demonstrated by monitoring the differential amino acids modification of native horse heart apomyoglobin/holomyoglobin and the human haptoglobin-hemoglobin complex. The obtained data confirmed the Togni reagent-mediated FFAP is an advantageous alternative method for covalent labeling in applications such as protein footprinting and epitope mapping of proteins (and their complexes) in general.

Název v anglickém jazyce

Fast Fluoroalkylation of Proteins Uncovers the Structure and Dynamics of Biological Macromolecules

Popis výsledku anglicky

Covalent labeling of proteins in combination with mass spectrometry has been established as a complementary technique to classical structural methods, such as X-ray, NMR, or cryogenic electron microscopy (Cryo-EM), used for protein structure determination. Although the current covalent labeling techniques enable the protein solvent accessible areas with sufficient spatial resolution to be monitored, there is still high demand for alternative, less complicated, and inexpensive approaches. Here, we introduce a new covalent labeling method based on fast fluoroalkylation of proteins (FFAP). FFAP uses fluoroalkyl radicals formed by reductive decomposition of Togni reagents with ascorbic acid to label proteins on a time scale of seconds. The feasibility of FFAP to effectively label proteins was demonstrated by monitoring the differential amino acids modification of native horse heart apomyoglobin/holomyoglobin and the human haptoglobin-hemoglobin complex. The obtained data confirmed the Togni reagent-mediated FFAP is an advantageous alternative method for covalent labeling in applications such as protein footprinting and epitope mapping of proteins (and their complexes) in general.

Druh

J_SC - Článek v periodiku v databázi SCOPUS

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/GA19-16084S" target="_blank" >GA19-16084S: Mapování přístupnosti proteinového povrchu kombinací hmotnostní spektrometrie a radikálového značení</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

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

Název periodika

Journal of the American Chemical Society

ISSN

0002-7863

e-ISSN

1520-5126

Svazek periodika

143

Číslo periodika v rámci svazku

49

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

10

Strana od-do

20670-20679

Kód UT WoS článku

000750799000004

EID výsledku v databázi Scopus

2-s2.0-85120891502