Electrospray ionization mass spectrometric characterization of acrylamide adducts to hemoglobin

Journal Article · Fri Jan 01 04:00:00 EST 1993 · Journal of Toxicology and Environmental Health; (United States)

DOI:https://doi.org/10.1080/15287399309531785· OSTI ID:7160744

Springer, D L; Goheen, S C; Edmonds, C G ^[1]; Bull, R J; Sylvester, D M ^[2]

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1. Pacific Northwest Lab., Richland, WA (United States)
2. Washington State Univ., Pullman, WA (United States)

The most common procedure to identify hemoglobin adducts has been to cleave the adducts from the protein and characterize the adducting species, by, for example, derivatization and gas chromatography/mass spectrometry. To extend these approaches we used electrospray ionization mass spectrometry (ESI-MS) to characterize adducted hemoglobin. For this we incubated [[sup 14]C]acrylamide with the purified human hemoglobin (type A[sub 0]) under conditions that yielded high adduct levels. When the hemoglobin was separated by reversed-phase high-performance liquid chromatography (HPLC), 65% of the radioactivity copurified with the [beta]-subunit. Three adducted species were prominent in the ESI mass spectrum of the intact [beta]-subunit, indicating acrylamide adduction (i.e., mass increase of 71 Da) and two addition unidentified moieties with mass increments of 102 and 135 Da. Endoproteinase Glu-C digestion of the adducted [beta]-subunit resulted in a peptide mixture that, upon reversed-phase HPLC separation, provided several radiolabeled peptides. Using ESI-MS we identified these as the V[sub 91-101] and V[sub 102-122] peptides that represent the cysteine-containing peptides of the [beta]-subunit. These results provide definitive information on acrylamide-modified human hemoglobin and demonstrate that ESI-MS provides valuable structure information on chemically adducted proteins. 30 refs., 9 figs., 3 tabs.

• OSTI ID: 7160744
• Journal Information: Journal of Toxicology and Environmental Health; (United States), Journal Name: Journal of Toxicology and Environmental Health; (United States) Vol. 40:2-3; ISSN 0098-4108; ISSN JTEHD6
• Country of Publication: United States
• Language: English

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ACRYLAMIDE
ADDUCTS
AMIDES
CARBOXYLIC ACIDS
CARCINOGENESIS
CHEMICAL ANALYSIS
CHROMATOGRAPHY
EVALUATION
GLOBINS
HEMOGLOBIN
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
IONIZATION
LIQUID COLUMN CHROMATOGRAPHY
MASS SPECTROSCOPY
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PATHOGENESIS
PIGMENTS
PORPHYRINS
PROTEINS
QUANTITATIVE CHEMICAL ANALYSIS
RADIOCHEMICAL ANALYSIS
SEPARATION PROCESSES
SPECTROSCOPY