The identification and partial characterization of acetaldehyde adducts of hemoglobin occurring in vivo: a possible marker of alcohol consumption.
Chromatographic, peptide mapping and mass spectrometric analysis were used to examine hemoglobin (Hb) from heavy drinkers and abstainers for alcohol consumption-related modifications. Heavy drinker and abstainer hemoglobin samples contained similar amounts of glycosylated Hb and significantly different (p < 0.05) amounts of "fast" hemoglobin. The presence of higher amounts of "fast" Hb in heavy drinker relative to abstainer samples suggested the presence of alcohol-consumption related modifications. To further examine Hb for modifications, tryptic peptides of the "fast" hemoglobin HbA1c were isolated and analyzed by plasma desorption mass spectrometry (PDMS). [14C]acetaldehyde (AcH)-Hb was synthesized in vivo for use as a standard. Specific peptides were chosen based on co-migration with radiolabeled peptides from a tryptic digest of the [14C]acetaldehyde-Hb. The masses obtained by PDMS for two heavy drinker peptides were identical to two radiolabeled peptides; the two pairs of peptides co-migrated on HPLC. A comparison of the observed mass for the peptides with the theoretical masses for acetaldehyde-modified Hb peptides suggested that the peptides were AcH-modified alpha and beta chain N-termini of Hb. The modified peptides were found in five of six heavy drinker samples. This is the first description of site-specific AcH-Hb adducts occurring in vivo. The routine detection of such adducts has potential for characterizing usual alcohol intake.
Gross
M D
MD
Division of Epidemiology, School of Public Health, University of Minnesota, Minneapolis 55454-1015.
Gapstur
S M
SM
Belcher
J D
JD
Scanlan
G
G
Potter
J D
JD
D000079:Acetaldehyde / Q000097:blood*; D000328:Adult; D000428:Alcohol Drinking / Q000097:blood*; D000437:Alcoholism / Q000097:blood / Q000175:diagnosis* / Q000534:rehabilitation; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000378:metabolism; D005260:Female; D006442:Glycated Hemoglobin / Q000378:metabolism; D006441:Hemoglobin A / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D010455:Peptides / Q000097:blood; D011485:Protein Binding / Q000502:physiology
D000079:Acetaldehyde; D006442:Glycated Hemoglobin; D006441:Hemoglobin A; D006454:Hemoglobins; D010455:Peptides