Covalent modification of lysine residues by allyl isothiocyanate in physiological conditions: Plausible transformation of isothiocyanate from thiol to amine

Toshiyuki Nakamura, Yoshichika Kawai, Noritoshi Kitamoto, Toshihiko Osawa, Yoji Kato

Research output: Contribution to journalArticle

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Abstract

We investigated the reactivity of allyl isothiocyanate (AITC) with amino groups under physiological conditions. First, the chemical reaction of AITC with bovine serum albumin (BSA) was investigated. When BSA was incubated with AITC in a phosphate buffer (pH 7.4), the loss of Lys residues was observed. Second, the Lys residue Nα-benzoyl-glycyl-L-lysine (BGK) was reacted with AITC in the buffer, and a novel peak was detected using high performance liquid chromatography (HPLC). The peak was purified and identified as AITC-modified BGK with a N£-thiocarbamoyl linkage. However, a thiol residue is known to be a predominant target of an isothiocyanate (ITC). Although AITC may react with a thiol moiety in vivo, a thiocarbamoyl linkage between ITC and thiol is unstable, and an AITC molecule may be regenerated. To prove the plausible transformation of ITC from thiol to amine, synthetic AITC-conjugated Nα-acetyl-L-cysteine (NAC) was incubated with BGK at 37 °C in physiological buffer, and the generation of AITC-Lys was analyzed. The loss of the AITC-NAC adduct corresponded to the formation of the AITC-BGK adduct. Furthermore, using a novel monoclonal antibody (A4C7mAb) specific for AITC-Lys, we found that the AITC-Lys residue was generated from the reaction between AITC-NAC and BSA. Although AITC preferentially reacts with thiol rather than with Lys, AITC can be liberated from thiols and can then react with amino groups. The ITC-Lys adduct may be a useful marker for ITC target molecules.

Original languageEnglish
Pages (from-to)536-542
Number of pages7
JournalChemical Research in Toxicology
Volume22
Issue number3
DOIs
Publication statusPublished - Mar 16 2009
Externally publishedYes

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Sulfhydryl Compounds
Lysine
Amines
Acetylcysteine
Bovine Serum Albumin
Buffers
allyl isothiocyanate
isothiocyanic acid
glycyllysine
Molecules
High performance liquid chromatography
Chemical reactions

ASJC Scopus subject areas

  • Toxicology

Cite this

Covalent modification of lysine residues by allyl isothiocyanate in physiological conditions : Plausible transformation of isothiocyanate from thiol to amine. / Nakamura, Toshiyuki; Kawai, Yoshichika; Kitamoto, Noritoshi; Osawa, Toshihiko; Kato, Yoji.

In: Chemical Research in Toxicology, Vol. 22, No. 3, 16.03.2009, p. 536-542.

Research output: Contribution to journalArticle

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T1 - Covalent modification of lysine residues by allyl isothiocyanate in physiological conditions

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AU - Osawa, Toshihiko

AU - Kato, Yoji

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