Application of Raney nickel to measure adducts of styrene oxide with hemoglobin and albumin.
Adducts of styrene 7,8-oxide (SO) with the blood proteins, Hb and albumin (Alb), were measured following treatment of the proteins with Raney nickel (Ra-Ni) to cleave 2-phenylethanol, which was subsequently assayed. Internal standards were prepared by modifying Hb and Alb with 4-methyl-SO to produce similar adducts. In a preliminary experiment with human blood, which had been modified with [14C]SO in vitro, it was determined that Ra-Ni released 6% of the total binding to globin and 76% of the total binding to Alb. Since Ra-Ni primarily cleaves covalently-bound sulfur species, this suggests that much more binding to human serum Alb occurred at a free cysteine residue than to human Hb. The overall rate of reaction of SO at 37 degrees C in vitro had a half-time of 0.70 h in human blood and of 0.36 h in blood from Sprague-Dawley rats. Adducts of SO with Hb and Alb were measured following SO modification of blood from humans and rats in vitro at 37 degrees C. Slopes of the linear relationships between adduct level and SO concentration allowed the following second-order rate constants [(L/mol of protein)/h] to be estimated: rat Hb, 72; human Hb, 2.4; rat Alb, 63; human Alb, 32. These rate constants show that intrinsic reactivity of SO toward the cysteine residue of human Hb is much lower than that of rat Hb or of Alb from either species. Adducts of Hb and Alb were also measured following administration of both SO and styrene to rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Rappaport
S M
SM
Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill 27599.
Ting
D
D
Jin
Z
Z
Yeowell-O'Connell
K
K
Waidyanatha
S
S
McDonald
T
T
D000818:Animals; D002849:Chromatography, Gas; D004852:Epoxy Compounds / Q000032:analysis*; D005260:Female; D005914:Globins / Q000032:analysis / Q000302:isolation & purification; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D008297:Male; D009532:Nickel / Q000737:chemistry*; D010626:Phenylethyl Alcohol / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D012044:Regression Analysis; D012709:Serum Albumin / Q000032:analysis*
D004852:Epoxy Compounds; D006454:Hemoglobins; D012709:Serum Albumin; D009532:Nickel; D005914:Globins; C013690:styrene oxide; D010626:Phenylethyl Alcohol