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Albumin as a carrier for NO delivery: preparation, physicochemical characterization, and interaction with gold nanoparticles

Abstract : BACKGROUND:Nitric oxide (NO) is a gaseous transmitter playing numerous physiological roles and characterized by a short half-life. Its binding to endogenous thiols increases its stability, facilitating its storage and transport. The purpose of this study was to investigate the nitrosated serum albumin (SA-SNO) and to provide a reference for its easy preparation for further use in in vitro studies.METHODS:Serum albumin (SA) was S-nitrosated by reacting with (i) NaNO2 in acidic medium; (ii) different low-molecular weight S-nitrosothiols (RSNO) (S-nitrosocysteine (CysNO), S-nitrosoglutathione (GSNO), and S,S'-dinitrosobucillamine (Buc(NO)2)); and (iii) diethylamine NONOate (DEA/NO). SA-SNO was purified by size exclusion chromatography and the S-nitrosation site and the rate were studied by mass spectrometry and Griess-Saville assay, respectively. Then, SA-SNO was characterized by spectrofluorimetry, dynamic light scattering, and circular dichroism. Finally, SA-SNO reactivity with citrate stabilized gold nanoparticles (AuNP-citrate) was investigated via determination of NO release.RESULTS:S-nitrosation rates of SA were 90.1 ± 3.3, 76.8 ± 2.7, 80.3 ± 3.2, 84.8 ± 5.0, and 15.4 ± 1.9% (n = 5), when SA was reacted with acidified NaNO2, CysNO, GSNO, Buc(NO)2, and DEA/NO, respectively. The physicochemical characterization indicated that the resulting product corresponded to a mono-S-nitrosothiol (on cysteine-34), and the conformational construction remained unchanged. Stability studies showed that the NO content was preserved over 1 week. AuNP-citrate reacted with SA-SNO with increase of its hydrodynamic diameter but preservation of SNO bond.CONCLUSIONS:SA-SNO prepared and stored under the reported conditions affords a well-defined reference suitable for in vitro studies.
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Submitted on : Thursday, October 13, 2016 - 9:26:37 AM
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Ming Luo, Ariane Boudier, Arnaud Pallotta, Philippe Maincent, Jean-Baptiste Vincourt, et al.. Albumin as a carrier for NO delivery: preparation, physicochemical characterization, and interaction with gold nanoparticles. Drug Development and Industrial Pharmacy, Taylor & Francis, 2016, 42 (12), pp.1928-1937. ⟨10.1080/03639045.2016.1182546⟩. ⟨hal-01380405⟩



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