Trimethylacetic Anhydride-Based Derivatization Facilitates
Quantification of Histone Marks at the MS1 Level

J 2021

Trimethylacetic Anhydride-Based Derivatization Facilitates Quantification of Histone Marks at the MS1 Level

KUCHAŘÍKOVÁ, Hana, Pavlína DOBROVOLNÁ, Gabriela LOCHMANOVÁ and Zbyněk ZDRÁHAL

Basic information

Original name

Trimethylacetic Anhydride-Based Derivatization Facilitates Quantification of Histone Marks at the MS1 Level

Authors

KUCHAŘÍKOVÁ, Hana (203 Czech Republic, belonging to the institution), Pavlína DOBROVOLNÁ (203 Czech Republic, belonging to the institution), Gabriela LOCHMANOVÁ (203 Czech Republic, belonging to the institution) and Zbyněk ZDRÁHAL (203 Czech Republic, guarantor, belonging to the institution)

Edition

Molecular and Cellurar Proteomic, American Society for Biochemistry and Molecular Biology, 2021, 1535-9476

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

fulltext

Impact factor

Impact factor: 5.911 in 2020

RIV identification code

RIV/00216224:14740/21:00119599

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1016/j.mcpro.2021.100114

UT WoS

000674937100001

Keywords in English

POSTTRANSLATIONAL MODIFICATIONS; DOWN PROTEOMICS; PROPIONYLATION; IDENTIFICATION; METHYLATION; ACETYLATION

Abstract

V originále

Histone post-translational modifications (hPTMs) are epigenetic marks that strongly affect numerous processes, including cell cycling and protein interactions. They have been studied by both antibody- and MS-based methods for years, but the analyses are still challenging, mainly because of the diversity of histones and their modifications arising from high contents of reactive amine groups in their amino acid sequences. Here, we introduce use of trimethylacetic anhydride (TMA) as a new reagent for efficient histone derivatization, which is a requirement for bottom-up proteomic hPTM analysis. TMA can denyatize unmodified amine groups of lysine residues and amine groups generated at peptide N-termini by trypsin digestion. The derivatization is facilitated by microwave irradiation, which also reduces incubation times to minutes. We demonstrate that histone derivatization with TMA reliably provides high yields of fully derivatized peptides and thus is an effective alternative to conventional methods. TMA afforded more than 98% and 99% labeling efficiencies for histones H4 and H3, respectively, thereby enabling accurate quantification of peptide forms. Trimethylacetylation substantially improves chromatographic separation of peptide forms, which is essential for direct quantification based on signals extracted from MS1 data. For this purpose, software widely applied by the proteomics community can be used without additional computational development. Thorough comparison with widely applied propionylation highlights the advantages of TMA-based histone derivatization for monitoring hPTMs in biological samples.

Links

GF19-29701L, research and development project

Name: Funkce HDAC1 v T-buněčných lymfomech

Investor: Czech Science Foundation, Partner Agency (Austria)

LM2018127, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

LM2018140, research and development project

Name: e-Infrastruktura CZ (Acronym: e-INFRA CZ)

Investor: Ministry of Education, Youth and Sports of the CR

90127, large research infrastructures

Name: CIISB II

Citovat

KUCHAŘÍKOVÁ, Hana, Pavlína DOBROVOLNÁ, Gabriela LOCHMANOVÁ and Zbyněk ZDRÁHAL. Trimethylacetic Anhydride-Based Derivatization Facilitates Quantification of Histone Marks at the MS1 Level. Molecular and Cellurar Proteomic. American Society for Biochemistry and Molecular Biology, 2021, vol. 20, JAN 01, p. "100114", 14 pp. ISSN 1535-9476. Available from: https://dx.doi.org/10.1016/j.mcpro.2021.100114.

@article{1824743,
   author = {Kuchaříková, Hana and Dobrovolná, Pavlína and Lochmanová, Gabriela and Zdráhal, Zbyněk},
   article_number = {JAN 01},
   doi = {http://dx.doi.org/10.1016/j.mcpro.2021.100114},
   keywords = {POSTTRANSLATIONAL MODIFICATIONS; DOWN PROTEOMICS; PROPIONYLATION; IDENTIFICATION; METHYLATION; ACETYLATION},
   language = {eng},
   issn = {1535-9476},
   journal = {Molecular and Cellurar Proteomic},
   title = {Trimethylacetic Anhydride-Based Derivatization Facilitates Quantification of Histone Marks at the MS1 Level},
   url = {https://www.mcponline.org/article/S1535-9476(21)00086-4/pdf},
   volume = {20},
   year = {2021}
}

TY  - JOUR
ID  - 1824743
AU  - Kuchaříková, Hana - Dobrovolná, Pavlína - Lochmanová, Gabriela - Zdráhal, Zbyněk
PY  - 2021
TI  - Trimethylacetic Anhydride-Based Derivatization Facilitates Quantification of Histone Marks at the MS1 Level
JF  - Molecular and Cellurar Proteomic
VL  - 20
IS  - JAN 01
SP  - "100114"
EP  - "100114"
PB  - American Society for Biochemistry and Molecular Biology
SN  - 15359476
KW  - POSTTRANSLATIONAL MODIFICATIONS
KW  - DOWN PROTEOMICS
KW  - PROPIONYLATION
KW  - IDENTIFICATION
KW  - METHYLATION
KW  - ACETYLATION
UR  - https://www.mcponline.org/article/S1535-9476(21)00086-4/pdf
N2  - Histone post-translational modifications (hPTMs) are epigenetic marks that strongly affect numerous processes, including cell cycling and protein interactions. They have been studied by both antibody- and MS-based methods for years, but the analyses are still challenging, mainly because of the diversity of histones and their modifications arising from high contents of reactive amine groups in their amino acid sequences. Here, we introduce use of trimethylacetic anhydride (TMA) as a new reagent for efficient histone derivatization, which is a requirement for bottom-up proteomic hPTM analysis. TMA can denyatize unmodified amine groups of lysine residues and amine groups generated at peptide N-termini by trypsin digestion. The derivatization is facilitated by microwave irradiation, which also reduces incubation times to minutes. We demonstrate that histone derivatization with TMA reliably provides high yields of fully derivatized peptides and thus is an effective alternative to conventional methods. TMA afforded more than 98% and 99% labeling efficiencies for histones H4 and H3, respectively, thereby enabling accurate quantification of peptide forms. Trimethylacetylation substantially improves chromatographic separation of peptide forms, which is essential for direct quantification based on signals extracted from MS1 data. For this purpose, software widely applied by the proteomics community can be used without additional computational development. Thorough comparison with widely applied propionylation highlights the advantages of TMA-based histone derivatization for monitoring hPTMs in biological samples.
ER  -

KUCHAŘÍKOVÁ, Hana, Pavlína DOBROVOLNÁ, Gabriela LOCHMANOVÁ and Zbyněk ZDRÁHAL. Trimethylacetic Anhydride-Based Derivatization Facilitates Quantification of Histone Marks at the MS1 Level. \textit{Molecular and Cellurar Proteomic}. American Society for Biochemistry and Molecular Biology, 2021, vol.~20, JAN 01, p.~''100114'', 14 pp. ISSN~1535-9476. Available from: https://dx.doi.org/10.1016/j.mcpro.2021.100114.

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