Adductomics Pipeline for Untargeted Analysis of Modifications to Cys34 of Human Serum Albumin.
An important but understudied class of human exposures is comprised of reactive electrophiles that cannot be measured in vivo because they are short-lived. An avenue for assessing these meaningful exposures focuses on adducts from reactions with nucleophilic loci of blood proteins, particularly Cys34 of human serum albumin, which is the dominant scavenger of reactive electrophiles in serum. We developed an untargeted analytical scheme and bioinformatics pipeline for detecting, quantitating, and annotating Cys34 adducts in tryptic digests of human serum/plasma. The pipeline interrogates tandem mass spectra to find signatures of the Cys34-containing peptide, obtains accurate masses of putative adducts, quantitates adduct levels relative to a "housekeeping peptide", and annotates modifications based on a combination of retention time, accurate mass, elemental composition, and database searches. We used the adductomics pipeline to characterize 43 adduct features in archived plasma from healthy human subjects and found several that were highly associated with smoking status, race, and other covariates. Since smoking is a strong risk factor for cancer and cardiovascular disease, our ability to discover adducts that distinguish smokers from nonsmokers with untargeted adductomics indicates that the pipeline is suitable for use in epidemiologic studies. In fact, adduct features were both positively and negatively associated with smoking, indicating that some adducts arise from reactions between Cys34 and constituents of cigarette smoke (e.g., ethylene oxide and acrylonitrile) while others (Cys34 oxidation products and disulfides) appear to reflect alterations in the serum redox state that resulted in reduced adduct levels in smokers.
School of Public Health, University of California , Berkeley, California 94720-7356, United States.
Hasmik
H
Grigoryan
School of Public Health, University of California , Berkeley, California 94720-7356, United States.
William
W
Edmands
School of Public Health, University of California , Berkeley, California 94720-7356, United States.
Sixin S
SS
Lu
School of Public Health, University of California , Berkeley, California 94720-7356, United States.
Yukiko
Y
Yano
School of Public Health, University of California , Berkeley, California 94720-7356, United States.
Luca
L
Regazzoni
California Institute for Quantitative Biosciences, University of California , Berkeley, California 94720, United States.
Anthony T
AT
Iavarone
College of Chemistry, University of California , Berkeley, California 94720, United States.
Evan R
ER
Williams
School of Public Health, University of California , Berkeley, California 94720-7356, United States.
Stephen M
SM
Rappaport
D000073865:Cigarette Smoking / Q000097:blood / Q000378:metabolism; D003545:Cysteine / Q000031:analogs & derivatives / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D000075462:Serum Albumin, Human / Q000737:chemistry* / Q000378:metabolism; D055815:Young Adult
D010455:Peptides; D003545:Cysteine; D000075462:Serum Albumin, Human
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