Gross M D, Gapstur S M, Belcher J D, Scanlan G, Potter J D
Division of Epidemiology, School of Public Health, University of Minnesota, Minneapolis 55454-1015.
Alcohol Clin Exp Res. 1992 Dec;16(6):1093-103. doi: 10.1111/j.1530-0277.1992.tb00704.x.
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.
采用色谱分析、肽图分析和质谱分析方法，检测酗酒者和戒酒者的血红蛋白（Hb），以寻找与酒精摄入相关的修饰。酗酒者和戒酒者的血红蛋白样本中糖化血红蛋白含量相似，而“快速”血红蛋白含量存在显著差异（p < 0.05）。与戒酒者样本相比，酗酒者样本中“快速”Hb含量更高，这表明存在与酒精摄入相关的修饰。为进一步检测Hb的修饰情况，分离了“快速”血红蛋白HbA1c的胰蛋白酶肽段，并通过等离子体解吸质谱（PDMS）进行分析。体内合成了[14C]乙醛（AcH）-Hb用作标准品。根据与[14C]乙醛-Hb胰蛋白酶消化产物中放射性标记肽段的共迁移情况选择特定肽段。PDMS测得的两个酗酒者肽段的质量与两个放射性标记肽段相同；这两对肽段在高效液相色谱（HPLC）上共迁移。将观察到的肽段质量与乙醛修饰的Hb肽段的理论质量进行比较，表明这些肽段是Hb的AcH修饰的α和β链N端。在六个酗酒者样本中的五个样本中发现了修饰肽段。这是首次对体内发生的位点特异性AcH-Hb加合物进行描述。常规检测此类加合物具有表征通常酒精摄入量的潜力。
