Bioconjugation with Aminoalkylhydrazine for Efficient Mass Spectrometry-Based Detection of Small Carbonyl Compounds

Senthil K. Thangaraj, Sanni Voutilainen, Martina Andberg, Anu Koivula, Janne Jänis, Juha Rouvinen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Bioconjugation through oxime or hydrazone formation is a versatile strategy for covalent labeling of biomolecules in vitro and in vivo. In this work, a mass spectrometry-based method was developed for the bioconjugation of small carbonyl compounds (CCs) with an aminoalkylhydrazine to form stable hydrazone conjugates that are readily detectable with electrospray ionization mass spectrometry (ESI-MS). Out of all hydrazine reagents tested, 2-(dimethylamino)ethylhydrazine (DMAEH) was selected for further analysis due to the fastest reaction rates observed. A thorough study of the reaction kinetics between structurally varied short-chain CCs and DMAEH was performed with the second-order reaction rate constants spanning in the range of 0.23-208 M-1 s-1. In general, small aldehydes reacted faster than the corresponding ketones. Moreover, a successful reaction monitoring of a deoxyribose-5-phosphate aldolase-catalyzed reversible retro-aldol cleavage of deoxyribose was demonstrated. Thus, the developed method shows potential also for ESI-MS-based enzyme kinetics studies.

Original languageEnglish
Pages (from-to)13447 - 13453
Number of pages7
JournalACS Omega
Volume4
Issue number8
DOIs
Publication statusPublished - 20 Aug 2019
MoE publication typeA1 Journal article-refereed

Fingerprint

Carbonyl compounds
Deoxyribose
Mass spectrometry
Hydrazones
Electrospray ionization
hydrazine
Reaction rates
Enzyme kinetics
Fructose-Bisphosphate Aldolase
Oximes
Hydrazine
Biomolecules
Ketones
Aldehydes
Reaction kinetics
Labeling
Rate constants
Phosphates
Monitoring
ethylhydrazine

Cite this

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title = "Bioconjugation with Aminoalkylhydrazine for Efficient Mass Spectrometry-Based Detection of Small Carbonyl Compounds",
abstract = "Bioconjugation through oxime or hydrazone formation is a versatile strategy for covalent labeling of biomolecules in vitro and in vivo. In this work, a mass spectrometry-based method was developed for the bioconjugation of small carbonyl compounds (CCs) with an aminoalkylhydrazine to form stable hydrazone conjugates that are readily detectable with electrospray ionization mass spectrometry (ESI-MS). Out of all hydrazine reagents tested, 2-(dimethylamino)ethylhydrazine (DMAEH) was selected for further analysis due to the fastest reaction rates observed. A thorough study of the reaction kinetics between structurally varied short-chain CCs and DMAEH was performed with the second-order reaction rate constants spanning in the range of 0.23-208 M-1 s-1. In general, small aldehydes reacted faster than the corresponding ketones. Moreover, a successful reaction monitoring of a deoxyribose-5-phosphate aldolase-catalyzed reversible retro-aldol cleavage of deoxyribose was demonstrated. Thus, the developed method shows potential also for ESI-MS-based enzyme kinetics studies.",
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Bioconjugation with Aminoalkylhydrazine for Efficient Mass Spectrometry-Based Detection of Small Carbonyl Compounds. / Thangaraj, Senthil K.; Voutilainen, Sanni; Andberg, Martina; Koivula, Anu; Jänis, Janne; Rouvinen, Juha (Corresponding Author).

In: ACS Omega, Vol. 4, No. 8, 20.08.2019, p. 13447 - 13453.

Research output: Contribution to journalArticleScientificpeer-review

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