title: An LC/ESI-SRM/MS method to screen chemically modified hemoglobin: simultaneous analysis for oxidized, nitrated, lipidated, and glycated sites.
abstract: Proteins are continuously exposed to various reactive chemical species (reactive oxygen/nitrogen species, endogenous/exogenous aldehydes/epoxides, etc.) due to physiological and chemical stresses, resulting in various chemical modifications such as oxidation, nitration, glycation/glycoxidation, lipidation/lipoxidation, and adduct formation with drugs/chemicals. Abundant proteins with a long half-life, such as hemoglobin (Hb, t 1/2 63 days, ∼150 mg/mL), are believed to be major targets of reactive chemical species that reflect biological events. Chemical modifications on Hb have been investigated mainly by mechanistic in vitro experiments or in vivo/clinical experiments focused on single target modifications. Here, we describe an optimized LC/ESI-SRM/MS method to screen oxidized, nitrated, lipidated, and glycated sites on Hb. In vivo preliminary results suggest that this method can detect simultaneously the presence of oxidation (+16 Da) of α-Met(32), α-Met(76), β-Met(55), and β-Trp(15) and adducts of malondialdehyde (+54 Da) and glycation (+162 Da) of β-Val(1) in a blood sample from a healthy volunteer. Graphical Abstract Screening chemical modifications on hemoglobin.
authors.affiliation: Department of Bioanalytical Chemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aramaki-aoba, Aoba-ku, Sendai, 980-8578, Japan.|Head Office/Research Laboratories, Ono Pharmaceutical Co., Ltd., 8-2 Kyutaromachi 1-chome, Chuo-ku, Osaka, 541-8564, Japan.
authors.forename: Koki
authors.initials: K
authors.lastname: Kojima
authors.affiliation: Department of Bioanalytical Chemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aramaki-aoba, Aoba-ku, Sendai, 980-8578, Japan.
authors.forename: Seon Hwa
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Department of Bioanalytical Chemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aramaki-aoba, Aoba-ku, Sendai, 980-8578, Japan. t-oe@mail.pharm.tohoku.ac.jp.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Oe
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D001665:Binding Sites; D001774:Blood Chemical Analysis / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D017127:Glycation End Products, Advanced / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry*; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D009566:Nitrates / Q000737:chemistry*; D010084:Oxidation-Reduction; D011485:Protein Binding; D017382:Reactive Oxygen Species / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000596:Amino Acids; D017127:Glycation End Products, Advanced; D006454:Hemoglobins; D008055:Lipids; D009566:Nitrates; D017382:Reactive Oxygen Species
keywords: Chemical modification; Hemoglobin; Mass spectrometry; Proteomics