Chemical methods for the detection of protein N-homocysteinylation via selective reactions with aldehydes

Tianzhu Zang, Shujia Dai, Dajun Chen, Bobby W.K. Lee, Suli Liu, Barry L. Karger, Zhaohui Sunny Zhou

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Elevated blood levels of homocysteine (Hcy), hyperhomocysteinemia or homocystinuria, have been associated with various diseases and conditions. Homocysteine thiolactone (Hcy TL) is a metabolite of Hcy and reacts with amine groups in proteins to form stable amides, homocystamides, or N-homocysteinylated proteins. It has been proposed that protein N-homocysteinylation contributes to the cytotoxicity of elevated Hcy. Due to its heterogeneity and relatively low abundance, detection of this posttranslational modification remains challenging. On the other hand, the γ-aminothiol group in homocystamides imparts different chemical reactivities than the native proteins. Under mildly acidic conditions, γ-aminothiols irreversibly and stoichiometrically react with aldehydes to form stable 1,3-thiazines, whereas the reversible Schiff base formation between aldehydes and amino groups in native proteins is markedly disfavored due to protonation of amines. As such, we have developed highly selective chemical methods to derivatize N-homocysteinylated proteins with various aldehyde tags, thereby facilitating the subsequent analyses. For instance, fluorescent or biotin tagging coupled with gel electrophoresis permits quantification and global profiling of complex biological samples, such as hemoglobin and plasma from rat, mouse and human; affinity enrichment with aldehyde resins drastically reduces sample complexity. In addition, different reactivities of lysine residues in hemoglobin toward Hcy TL were observed.

Original languageEnglish (US)
Pages (from-to)9065-9071
Number of pages7
JournalAnalytical Chemistry
Volume81
Issue number21
DOIs
StatePublished - Nov 1 2009
Externally publishedYes

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Aldehydes
Homocysteine
Proteins
Amines
Hemoglobins
Thiazines
Chemical reactivity
Schiff Bases
Protonation
Cytotoxicity
Biotin
Metabolites
Electrophoresis
Amides
Lysine
Rats
Blood
Resins
Gels
Plasmas

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Chemical methods for the detection of protein N-homocysteinylation via selective reactions with aldehydes. / Zang, Tianzhu; Dai, Shujia; Chen, Dajun; Lee, Bobby W.K.; Liu, Suli; Karger, Barry L.; Zhou, Zhaohui Sunny.

In: Analytical Chemistry, Vol. 81, No. 21, 01.11.2009, p. 9065-9071.

Research output: Contribution to journalArticle

Zang, Tianzhu ; Dai, Shujia ; Chen, Dajun ; Lee, Bobby W.K. ; Liu, Suli ; Karger, Barry L. ; Zhou, Zhaohui Sunny. / Chemical methods for the detection of protein N-homocysteinylation via selective reactions with aldehydes. In: Analytical Chemistry. 2009 ; Vol. 81, No. 21. pp. 9065-9071.
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