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The interplay between copper(II), human serum albumin, fatty acids, and carbonylating agent interferes with Cys 34 thiol reactivity and copper binding

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Abstract

Cys34 thiol group of human serum albumin (HSA) represents major plasma antioxidant. Its reactivity is influenced by multiple factors. The influence of fatty acids (FA; saturated, mono, and poly unsaturated acids from fish oil) binding to HSA, on copper(II) binding affinity and Cys34 thiol group accessibility/reactivity, in the presence of carbonylation agent (methylglyoxal, MG) was examined. HSA–copper(II) content, thiol group reactivity, and HSA carbonylation level were monitored spectrophotometrically. Changes in HSA were followed by fluorescence spectroscopy and native PAG electrophoresis. FA/HSA molar ratio was screened by GC. Together, binding of copper(II) ions and FA to HSA increase the reactivity of Cys34 thiol group (depending on the type of FA), with constant contribution of copper(II) ions of one-third. Carbonylation of FA–HSA–Cu(II) complexes caused a decrease in the Cys34 thiol group content, accompanied by a decrease in the content of HSA-bound copper. The carbonylation level of guanidine groups was not affected by FAs and copper(II) binding. Fluorescent emission spectra of FA–HSA–Cu(II)–MG complexes showed conformational changes in HSA molecule. Although binding of fatty acids and copper ions caused a significant increase in the thiol group reactivity, Cys34 thiol from FA–HSA–Cu(II) complexes reacted with MG in smaller extent than expected, probably as a consequence of conformational changes introduced by carbonylation. Increase in the percentage of reacted-free thiol groups with MG (due to FA and copper binding) may not seem to be very significant, but it is very important in complex biological systems, where catalytic metal is present.

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Abbreviations

FA:

Fatty acids

Myr:

Myristic acid

Ole:

Oleic acid

Ste:

Stearic acid

FO:

Fish oil fatty acids

HSA:

Human serum albumin

Def HSA:

Defatted HSA

FA–HSA–Cu(II):

Complex HSA with FA and copper(II) ion

HSA–SH:

Cys34 free thiol group of HSA

MG:

Methylglyoxal

HSA–MG:

HSA modified with MG

DTNB:

(5,5′dithiobis-(2-nitrobenzoic acid)

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Acknowledgements

This work was supported by The Ministry of Education, Science and Technological Development of Serbia with Grant no. 172049. The authors acknowledge support of the FP7 RegPot Project FCUB ERA GA no. 256716.

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Authors and Affiliations

  1. Faculty of Chemistry, University of Belgrade, Studentski trg 12-16, 11000, Belgrade, Serbia

    Jelena M. Aćimović, Ivan D. Pavićević, Vesna B. Jovanović & Ljuba M. Mandić

  2. Institute for the Application of Nuclear Energy, INEP, Banatska 31b, 11000, Belgrade, Serbia

    Ana Z. Penezić

  3. Laboratory for Nutrition and Metabolism, Institute for Medical Research, University of Belgrade, Belgrade, Serbia

    Marija Takić

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  1. Ana Z. Penezić
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  2. Jelena M. Aćimović
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  3. Ivan D. Pavićević
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  6. Ljuba M. Mandić
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Correspondence to Ana Z. Penezić.

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Penezić, A.Z., Aćimović, J.M., Pavićević, I.D. et al. The interplay between copper(II), human serum albumin, fatty acids, and carbonylating agent interferes with Cys 34 thiol reactivity and copper binding. J Biol Inorg Chem 24, 61–70 (2019). https://doi.org/10.1007/s00775-018-1628-7

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  • DOI: https://doi.org/10.1007/s00775-018-1628-7

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