Albumin Thiolate Anion Is an Intermediate in the Formation of Albumin-S-S-Homocysteine

Shantanu Sengupta, Hong Chen, Tadayasu Togawa, Patricia M. DiBello, Alana K. Majors, Beatrix Büdy, Michael E Ketterer, Donald W. Jacobsen

Research output: Contribution to journalArticle

140 Citations (Scopus)

Abstract

An elevated concentration of plasma total homocysteine is an independent risk factor for cardiovascular disease. Greater than 80% of circulating homocysteine is covalently bound to plasma protein by disulfide bonds. It is known that albumin combines with cysteine in circulation to form albumin-Cys34-S-S-Cys. Studies are now presented to show that the formation of albumin-bound homocysteine proceeds through the generation of an albumin thiolate anion. Incubation of human plasma with L- 35S-homocysteine results in the association of >90% of the protein-bound 35S-homocysteine with albumin as shown by nonreduced SDS-polyacrylamide gel electrophoresis. Treatment of the complex with β-mercaptoethanol results in near quantitative release of the bound L- 35S-homocysteine, demonstrating that the binding of homocysteine to albumin is through a disulfide bond. Furthermore, using an in vitro model system to study the mechanisms of this disulfide bond formation, we show that homocysteine binds to albumin in two steps. In the first step homocysteine rapidly displaces cysteine from albumin-Cys34-S-S-Cys, forming albumin-Cys34 thiolate anion and homocysteine-cysteine mixed disulfide. In the second step, albumin thiolate anion attacks homocysteine-cysteine mixed disulfide to yield primarily albumin-Cys 34-S-S-Hcy and to a much lesser extent albumin-Cys 34-S-S-Cys. The results clearly suggest that when reduced homocysteine enters circulation, it attacks albumin-Cys34-S-S-Cys to form albumin-Cys34 thiolate anion, which in turn, reacts with homocysteine-cysteine mixed disulfide or homocystine to form albumin-bound homocysteine.

Original languageEnglish (US)
Pages (from-to)30111-30117
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number32
DOIs
StatePublished - Aug 10 2001

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Anions
Albumins
Homocysteine
Disulfides
albumin-S-S-homocysteine
Cysteine
Homocystine
Plasma (human)
Mercaptoethanol
Electrophoresis
Blood Proteins
Polyacrylamide Gel Electrophoresis
Cardiovascular Diseases
Association reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sengupta, S., Chen, H., Togawa, T., DiBello, P. M., Majors, A. K., Büdy, B., ... Jacobsen, D. W. (2001). Albumin Thiolate Anion Is an Intermediate in the Formation of Albumin-S-S-Homocysteine. Journal of Biological Chemistry, 276(32), 30111-30117. https://doi.org/10.1074/jbc.M104324200

Albumin Thiolate Anion Is an Intermediate in the Formation of Albumin-S-S-Homocysteine. / Sengupta, Shantanu; Chen, Hong; Togawa, Tadayasu; DiBello, Patricia M.; Majors, Alana K.; Büdy, Beatrix; Ketterer, Michael E; Jacobsen, Donald W.

In: Journal of Biological Chemistry, Vol. 276, No. 32, 10.08.2001, p. 30111-30117.

Research output: Contribution to journalArticle

Sengupta, S, Chen, H, Togawa, T, DiBello, PM, Majors, AK, Büdy, B, Ketterer, ME & Jacobsen, DW 2001, 'Albumin Thiolate Anion Is an Intermediate in the Formation of Albumin-S-S-Homocysteine', Journal of Biological Chemistry, vol. 276, no. 32, pp. 30111-30117. https://doi.org/10.1074/jbc.M104324200
Sengupta S, Chen H, Togawa T, DiBello PM, Majors AK, Büdy B et al. Albumin Thiolate Anion Is an Intermediate in the Formation of Albumin-S-S-Homocysteine. Journal of Biological Chemistry. 2001 Aug 10;276(32):30111-30117. https://doi.org/10.1074/jbc.M104324200
Sengupta, Shantanu ; Chen, Hong ; Togawa, Tadayasu ; DiBello, Patricia M. ; Majors, Alana K. ; Büdy, Beatrix ; Ketterer, Michael E ; Jacobsen, Donald W. / Albumin Thiolate Anion Is an Intermediate in the Formation of Albumin-S-S-Homocysteine. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 32. pp. 30111-30117.
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