title: Characterization of sulfur mustard induced structural modifications in human hemoglobin by liquid chromatography--tandem mass spectrometry.
abstract: In this paper we describe the use of tandem mass spectrometry to identify modified sites in human hemoglobin after in vitro exposure to bis(2-chloroethyl) sulfide (sulfur mustard). Globin isolated from human whole blood which had been exposed to sulfur mustard was degraded with trypsin, and the digests were analyzed by micro LC/MS. Alkylated tryptic fragments (alpha-T1, alpha-T4, alpha-T6, alpha-T9, beta-T1, beta-T9, beta-T10, beta-T11, and beta-T10-S-S-beta-T12) could be tentatively assigned upon comparison with a digest from nonexposed globin. Subsequent tandem mass spectrometry of these peptides allowed unambiguous assignment of 5 specific modified residues: alpha-Val-1, alpha-His-20, beta-Val-1, beta-His-77, and beta-His-97. The results demonstrate the usefulness of microbore LC in combination with tandem mass spectrometry for the structural determination of chemically modified peptides and proteins.
authors.affiliation: Department of Chemical Toxicology, TNO Prins Maurits Laboratory, Rijswijk, The Netherlands.
authors.forename: D
authors.initials: D
authors.lastname: Noort
authors.forename: E R
authors.initials: ER
authors.lastname: Verheij
authors.forename: A G
authors.initials: AG
authors.lastname: Hulst
authors.forename: L P
authors.initials: LP
authors.lastname: de Jong
authors.forename: H P
authors.initials: HP
authors.lastname: Benschop
mesh_terms: D000478:Alkylation; D000595:Amino Acid Sequence; D002273:Carcinogens / Q000032:analysis / Q000737:chemistry / Q000633:toxicity*; D002619:Chemical Warfare Agents / Q000032:analysis / Q000737:chemistry / Q000633:toxicity*; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000737:chemistry / Q000187:drug effects*; D008401:Gas Chromatography-Mass Spectrometry; D006454:Hemoglobins / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009151:Mustard Gas / Q000032:analysis / Q000737:chemistry / Q000633:toxicity*; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013462:Sulfur Radioisotopes; D014357:Trypsin / Q000378:metabolism
chemical_list: D002273:Carcinogens; D002619:Chemical Warfare Agents; D006454:Hemoglobins; D010446:Peptide Fragments; D013462:Sulfur Radioisotopes; D014357:Trypsin; D009151:Mustard Gas