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

    6 Citations (Scopus)

    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
    JournalACS Omega
    Volume4
    Issue number8
    DOIs
    Publication statusPublished - 20 Aug 2019
    MoE publication typeA1 Journal article-refereed

    Funding

    The FT-ICR facility is supported by Biocenter Finland, Biocenter Kuopio and the European Regional Development Fund (project number A70135). We thank Arja Kiema and Kirsi Kiiveri for technical assistance (VTT). This work received support from the Academy of Finland through SA-ENGBIOCAT project (Decision Numbers 287241 and 288677).

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