Plasma proteins as early biomarkers of exposure to carcinogenic aromatic amines.
Two-dimensional gel electrophoresis (2DG) has been used to study the changes induced in dog plasma polypeptides by the known urinary bladder carcinogens, 4-aminobiphenyl (4-ABP) and 2-naphthylamine (2-NA). Treatment with 3-aminobiphenyl (3-ABP) and 1-naphthylamine (1-NA), both considered to be non-carcinogenic, were used as controls. The purpose of this study was: (1) to determine whether or not changes that occurred in the plasma protein patterns were specific to 4-ABP and/or other related carcinogenic arylamines; (2) to measure the time course in the changes of the major polypeptides during dosing and their resynthesis during a recovery period; and (3) to determine, by microsequencing, the biochemical identity of the affected proteins. The results indicate that only the most potent carcinogen, 4-ABP, had the effect of suppressing the expression of some proteins, while the other aromatic amines caused no discernible change in the 2DG patterns during a 12-week dosing period. The 4-ABP caused dramatic suppression of two sets of proteins. One set of three spots had an apparent molecular weight of 32.5 kDa, and a pI of 5.8-6.0. The major component in this group was identified as the beta-chain of haptoglobin. Expression of this protein decreased markedly during the first 2 weeks of treatment and recovered slowly after dosing stopped. Since haptoglobin functions to bind with free hemoglobin and facilitates its elimination from the blood stream, these results can be rationalized as a consequence of 4-ABP binding to hemoglobin in the erythrocyte, resulting in cell death and hemolysis. The 4-ABP modified hemoglobin then binds to haptoglobin and this tertiary complex is purged from the blood stream, resulting in the disappearance of free haptoglobin. A second set of spots (mol. wt., 65 kDa; pI, 6.5-6.6) disappeared much faster than the haptoglobin, and recovered more quickly. The major protein is about one-fifth the intensity of haptoglobin and appeared to be N-terminally blocked. Internal microsequencing of four fragments obtained from tryptic cleavage of the major spot of this group showed significant similarity to the serum albumin sequence of several species. This spot group is not the major serum albumin spot, however, since the latter is readily identified as the most abundant spot on the plasma map. During the course of this study, several other polypeptides in the 2DG map of dog plasma were identified and are presented here.
Miller
M J
MJ
Laboratory of Experimental Carcinogenesis, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4255.
Parmelee
D C
DC
Benjamin
T
T
Sechi
S
S
Dooley
K L
KL
Kadlubar
F F
FF
D015081:2-Naphthylamine / Q000008:administration & dosage / Q000633:toxicity*; D000588:Amines / Q000633:toxicity*; D000595:Amino Acid Sequence; D000611:Aminobiphenyl Compounds / Q000008:administration & dosage / Q000633:toxicity*; D000818:Animals; D016632:Apolipoprotein A-I / Q000097:blood; D015415:Biomarkers; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D002273:Carcinogens / Q000633:toxicity*; D003198:Computer Simulation; D004285:Dogs; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005340:Fibrinogen / Q000378:metabolism; D006242:Haptoglobins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism
D000588:Amines; D000611:Aminobiphenyl Compounds; D016632:Apolipoprotein A-I; D015415:Biomarkers; D001798:Blood Proteins; D002273:Carcinogens; D006242:Haptoglobins; D012709:Serum Albumin; C006757:4-biphenylamine; D005340:Fibrinogen; D015081:2-Naphthylamine