title: Albumin is the main nucleophilic target of human plasma: a protective role against pro-atherogenic electrophilic reactive carbonyl species?
abstract: The aim of this work was to study the metabolic fate of 4-hydroxy- trans-2-nonenal (HNE) in human plasma, which represents the main vascular site of reactive carbonyl species (RCS) formation and where the main pro-atherogenic target proteins are formed. When HNE was spiked in human plasma, it rapidly disappeared (within 40 s) and no phase I metabolites were detected, suggesting that the main fate of HNE is due to an adduction mechanism. HNE consumption was then monitored in two plasma fractions: low molecular weight plasma protein fractions (<10 kDa; LMWF) and high molecular weight plasma protein fractions (>10 kDa; HMWF). HNE was almost stable in LMWF, while in HMWF it was consumed by almost 70% within 5 min. Proteomics identified albumin (HSA) as the main protein target, as further confirmed by a significantly reduced HNE quenching of dealbuminated plasma. LC-ESI-MS/MS analysis identified Cys34 and Lys199 as the most reactive adduction sites of HSA, through the formation of a Michael and Schiff base adducts, respectively. The rate constant of HNE trapping by albumin was 50.61 +/- 1.89 M (-1) s (-1) and that of Cys34 (29.37 M (-1) s (-1)) was 1 order of magnitude higher with respect to that of GSH (3.81 +/- 0.17 M (-1) s (-1)), as explained by molecular modeling studies. In conclusion, we suggest that albumin, through nucleophilic residues, and in particular Cys34, can act as an endogenous detoxifying agent of circulating RCS.
authors.affiliation: Istituto di Chimica Farmaceutica e Tossicologica Pietro Pratesi, Faculty of Pharmacy, University of Milan, I-20131, Milan, Italy. giancarlo.aldini@unimi.it
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Aldini
authors.forename: Giulio
authors.initials: G
authors.lastname: Vistoli
authors.forename: Luca
authors.initials: L
authors.lastname: Regazzoni
authors.forename: Luca
authors.initials: L
authors.lastname: Gamberoni
authors.forename: Roberto Maffei
authors.initials: RM
authors.lastname: Facino
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamaguchi
authors.forename: Koji
authors.initials: K
authors.lastname: Uchida
authors.forename: Marina
authors.initials: M
authors.lastname: Carini
mesh_terms: D000328:Adult; D000447:Aldehydes / Q000378:metabolism*; D050197:Atherosclerosis; D003545:Cysteine / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D020011:Protective Agents / Q000378:metabolism*; D012709:Serum Albumin / Q000378:metabolism*
chemical_list: D000447:Aldehydes; D010455:Peptides; D020011:Protective Agents; D012709:Serum Albumin; C027576:4-hydroxy-2-nonenal; D008239:Lysine; D003545:Cysteine