TY - JOUR
T1 - Alternative LC/ESI-MS/MS approach to screen hemoglobin N-terminal modifications
AU - Usuzawa, Go
AU - Lee, Seon Hwa
AU - Oe, Tomoyuki
N1 - Funding Information:
This work was supported in part by a Grant-in-Aid for Challenging Exploratory Research (to T.O., No. 15K14935 for 2015–2016) from the Japan Society for the Promotion of Science . The authors are indebted to Professor Ian A. Blair (University of Pennsylvania, Philadelphia, PA) and Professor Junken Aoki (The University of Tokyo, Tokyo, Japan) for donating a used LCQ Deca and TSQ Quantum Ultra, respectively.
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2021/10
Y1 - 2021/10
N2 - Proteins are continuously exposed to reactive chemical species owing to physiological and chemical stresses, resulting in various chemical modifications such as oxidation, nitration, glycation/glycoxidation, lipidation/lipoxidation, and adduct formation with drugs/chemicals. Hemoglobin (Hb) is the most abundant protein in blood (∼150 mg/mL) with a long half-life (turnover: 126 days). Hb is thus believed to be a major target of reactive chemical species and those modifications reflect biological events. Chemical modifications on Hb have been analyzed as N-terminal valine (Val) adducts by the Edman degradation reaction because Val is the N-terminal amino acid in both the α- and β-subunits of Hb and is exposed on the surface of the protein. However, this strategy is limited to N-terminal alkylated Val and overlooks N-terminal acylated and deaminated Val. Here, we describe a proof-of-concept study to develop an alternative screening approach based on liquid chromatography-tandem mass spectrometry with peptide-specific constant neutral loss/precursor ion scanning to include all types of N-terminal modifications on Hb.
AB - Proteins are continuously exposed to reactive chemical species owing to physiological and chemical stresses, resulting in various chemical modifications such as oxidation, nitration, glycation/glycoxidation, lipidation/lipoxidation, and adduct formation with drugs/chemicals. Hemoglobin (Hb) is the most abundant protein in blood (∼150 mg/mL) with a long half-life (turnover: 126 days). Hb is thus believed to be a major target of reactive chemical species and those modifications reflect biological events. Chemical modifications on Hb have been analyzed as N-terminal valine (Val) adducts by the Edman degradation reaction because Val is the N-terminal amino acid in both the α- and β-subunits of Hb and is exposed on the surface of the protein. However, this strategy is limited to N-terminal alkylated Val and overlooks N-terminal acylated and deaminated Val. Here, we describe a proof-of-concept study to develop an alternative screening approach based on liquid chromatography-tandem mass spectrometry with peptide-specific constant neutral loss/precursor ion scanning to include all types of N-terminal modifications on Hb.
KW - Chemical modifications
KW - Hemoglobin
KW - Mass spectrometry
KW - N-Terminal modifications
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U2 - 10.1016/j.ijms.2021.116651
DO - 10.1016/j.ijms.2021.116651
M3 - Article
AN - SCOPUS:85107728648
SN - 1387-3806
VL - 468
JO - International Journal of Mass Spectrometry and Ion Processes
JF - International Journal of Mass Spectrometry and Ion Processes
M1 - 116651
ER -