The formation of stable acetaldehyde-hemoglobin adducts in a red blood cell model

Susan M. Gapstur; Eugene G. DeMaster; John D. Potter; John D Belcher; Myron D Gross

doi:10.1016/0741-8329(92)90098-U

The formation of stable acetaldehyde-hemoglobin adducts in a red blood cell model

Susan M. Gapstur, Eugene G. DeMaster, John D. Potter, John D Belcher, Myron D Gross

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

The formation of stable hemoglobin adducts was examined (in the absence of an added reducing agent) in metabolizing red blood cells (RBCs) exposed to micromolar concentrations of acetaldehyde for up to 48 hours in vitro. The rapid disappearance of acetaldehyde due to oxidation by RBC aldehyde dehydrogenase was prevented by pretreating the cells with the inhibitor cyanamide. The RBCs remained viable for 48 hours (37°C) as determined by cell hemolysis and glycolytic activity. [¹⁴C]acetaldehyde-modified hemoglobin was assessed in untreated and in cyanamide-pretreated cells. In untreated cells, after 3 hours of exposure to 50 and 200 nmol/ml of [¹⁴C]acetaldehyde, the molar ratios of acetaldehyde to hemoglobin were 0.00069 and 0.0038, respectively; [¹⁴C]acetaldehyde concentrations decreased to less than 4% of the initial levels within 3 hours. In cyanamide-pretreated RBCs, the molar ratios of acetaldehyde bound to hemoglobin ranged from 0.0013 after 3 hours of exposure to 20 nmol/ml [¹⁴C]acetaldehyde up to 0.039 after 48 hours of exposure to 200 nmol/ml [¹⁴C]acetaldehyde. Following tryptic digestion of [¹⁴C]acetaldehyde-hemoglobin and separation of peptides by high-performance liquid chromatography, significant incorporation of [¹⁴C]acetaldehyde was observed in nine peptides. Modifications of the labeled peptides remain to be characterized.

Original language	English (US)
Pages (from-to)	563-569
Number of pages	7
Journal	Alcohol
Volume	9
Issue number	6
DOIs	https://doi.org/10.1016/0741-8329(92)90098-U
State	Published - 1992

Bibliographical note

Funding Information:
ACKNOWLEDGEMENTS This work was supported by NIAAA-NIH Research Grant ROI AA07838 to Dr. John Belcher and by the Department of Veteran Affairs. Ms. Gapstur was supported by NIH Training Grant T32 CA099607 to Dr. Potter. We are indebted to Thomas P. Krick for the GC/MS spectra. The GC/MS facilities were provided by the Biochemistry Department of the University of Minnesota and maintained by the Minnesota Agricultural Experiment Station. We thank Rebecca Hays for her excellent technical assistance.

Keywords

Acetaldehyde
Acetaldehyde-hemoglobin adducts
Acetaldehyde-modified hemoglobin
Cyanamide
Ethanol
Hemoglobin
Red blood cells

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10.1016/0741-8329(92)90098-U

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@article{d7e8739ca5614991be417672d503d4d3,

title = "The formation of stable acetaldehyde-hemoglobin adducts in a red blood cell model",

abstract = "The formation of stable hemoglobin adducts was examined (in the absence of an added reducing agent) in metabolizing red blood cells (RBCs) exposed to micromolar concentrations of acetaldehyde for up to 48 hours in vitro. The rapid disappearance of acetaldehyde due to oxidation by RBC aldehyde dehydrogenase was prevented by pretreating the cells with the inhibitor cyanamide. The RBCs remained viable for 48 hours (37°C) as determined by cell hemolysis and glycolytic activity. [14C]acetaldehyde-modified hemoglobin was assessed in untreated and in cyanamide-pretreated cells. In untreated cells, after 3 hours of exposure to 50 and 200 nmol/ml of [14C]acetaldehyde, the molar ratios of acetaldehyde to hemoglobin were 0.00069 and 0.0038, respectively; [14C]acetaldehyde concentrations decreased to less than 4% of the initial levels within 3 hours. In cyanamide-pretreated RBCs, the molar ratios of acetaldehyde bound to hemoglobin ranged from 0.0013 after 3 hours of exposure to 20 nmol/ml [14C]acetaldehyde up to 0.039 after 48 hours of exposure to 200 nmol/ml [14C]acetaldehyde. Following tryptic digestion of [14C]acetaldehyde-hemoglobin and separation of peptides by high-performance liquid chromatography, significant incorporation of [14C]acetaldehyde was observed in nine peptides. Modifications of the labeled peptides remain to be characterized.",

keywords = "Acetaldehyde, Acetaldehyde-hemoglobin adducts, Acetaldehyde-modified hemoglobin, Cyanamide, Ethanol, Hemoglobin, Red blood cells",

author = "Gapstur, {Susan M.} and DeMaster, {Eugene G.} and Potter, {John D.} and Belcher, {John D} and Gross, {Myron D}",

note = "Funding Information: ACKNOWLEDGEMENTS This work was supported by NIAAA-NIH Research Grant ROI AA07838 to Dr. John Belcher and by the Department of Veteran Affairs. Ms. Gapstur was supported by NIH Training Grant T32 CA099607 to Dr. Potter. We are indebted to Thomas P. Krick for the GC/MS spectra. The GC/MS facilities were provided by the Biochemistry Department of the University of Minnesota and maintained by the Minnesota Agricultural Experiment Station. We thank Rebecca Hays for her excellent technical assistance.",

year = "1992",

doi = "10.1016/0741-8329(92)90098-U",

language = "English (US)",

volume = "9",

pages = "563--569",

journal = "Alcohol",

issn = "0741-8329",

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T1 - The formation of stable acetaldehyde-hemoglobin adducts in a red blood cell model

AU - Gapstur, Susan M.

AU - DeMaster, Eugene G.

AU - Potter, John D.

AU - Belcher, John D

AU - Gross, Myron D

N1 - Funding Information: ACKNOWLEDGEMENTS This work was supported by NIAAA-NIH Research Grant ROI AA07838 to Dr. John Belcher and by the Department of Veteran Affairs. Ms. Gapstur was supported by NIH Training Grant T32 CA099607 to Dr. Potter. We are indebted to Thomas P. Krick for the GC/MS spectra. The GC/MS facilities were provided by the Biochemistry Department of the University of Minnesota and maintained by the Minnesota Agricultural Experiment Station. We thank Rebecca Hays for her excellent technical assistance.

PY - 1992

Y1 - 1992

N2 - The formation of stable hemoglobin adducts was examined (in the absence of an added reducing agent) in metabolizing red blood cells (RBCs) exposed to micromolar concentrations of acetaldehyde for up to 48 hours in vitro. The rapid disappearance of acetaldehyde due to oxidation by RBC aldehyde dehydrogenase was prevented by pretreating the cells with the inhibitor cyanamide. The RBCs remained viable for 48 hours (37°C) as determined by cell hemolysis and glycolytic activity. [14C]acetaldehyde-modified hemoglobin was assessed in untreated and in cyanamide-pretreated cells. In untreated cells, after 3 hours of exposure to 50 and 200 nmol/ml of [14C]acetaldehyde, the molar ratios of acetaldehyde to hemoglobin were 0.00069 and 0.0038, respectively; [14C]acetaldehyde concentrations decreased to less than 4% of the initial levels within 3 hours. In cyanamide-pretreated RBCs, the molar ratios of acetaldehyde bound to hemoglobin ranged from 0.0013 after 3 hours of exposure to 20 nmol/ml [14C]acetaldehyde up to 0.039 after 48 hours of exposure to 200 nmol/ml [14C]acetaldehyde. Following tryptic digestion of [14C]acetaldehyde-hemoglobin and separation of peptides by high-performance liquid chromatography, significant incorporation of [14C]acetaldehyde was observed in nine peptides. Modifications of the labeled peptides remain to be characterized.

AB - The formation of stable hemoglobin adducts was examined (in the absence of an added reducing agent) in metabolizing red blood cells (RBCs) exposed to micromolar concentrations of acetaldehyde for up to 48 hours in vitro. The rapid disappearance of acetaldehyde due to oxidation by RBC aldehyde dehydrogenase was prevented by pretreating the cells with the inhibitor cyanamide. The RBCs remained viable for 48 hours (37°C) as determined by cell hemolysis and glycolytic activity. [14C]acetaldehyde-modified hemoglobin was assessed in untreated and in cyanamide-pretreated cells. In untreated cells, after 3 hours of exposure to 50 and 200 nmol/ml of [14C]acetaldehyde, the molar ratios of acetaldehyde to hemoglobin were 0.00069 and 0.0038, respectively; [14C]acetaldehyde concentrations decreased to less than 4% of the initial levels within 3 hours. In cyanamide-pretreated RBCs, the molar ratios of acetaldehyde bound to hemoglobin ranged from 0.0013 after 3 hours of exposure to 20 nmol/ml [14C]acetaldehyde up to 0.039 after 48 hours of exposure to 200 nmol/ml [14C]acetaldehyde. Following tryptic digestion of [14C]acetaldehyde-hemoglobin and separation of peptides by high-performance liquid chromatography, significant incorporation of [14C]acetaldehyde was observed in nine peptides. Modifications of the labeled peptides remain to be characterized.

KW - Acetaldehyde

KW - Acetaldehyde-hemoglobin adducts

KW - Acetaldehyde-modified hemoglobin

KW - Cyanamide

KW - Ethanol

KW - Hemoglobin

KW - Red blood cells

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U2 - 10.1016/0741-8329(92)90098-U

DO - 10.1016/0741-8329(92)90098-U

M3 - Article

C2 - 1472315

AN - SCOPUS:0026442454

SN - 0741-8329

VL - 9

SP - 563

EP - 569

JO - Alcohol

JF - Alcohol

IS - 6

ER -

The formation of stable acetaldehyde-hemoglobin adducts in a red blood cell model

Abstract

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