Alpha 2u-globulin is the only member of the lipocalin protein superfamily that binds to hyaline droplet inducing agents.
The rate-limiting step in chemically induced, male rat-specific hyaline droplet nephropathy is the reversible binding of a xenobiotic to alpha 2u-globulin. In this study, equilibrium saturation binding experiments were conducted to evaluate the in vitro binding of d-limonene-1,2-oxide (dLO) and 2,4,4-trimethyl-2-pentanol (TMP-OH) to alpha 2u-globulin and members of the alpha 2u-globulin protein superfamily. Both dLO and TMP-OH bound to alpha 2u-globulin, with Scatchard analysis yielding dissociation constants of 5.6 and 6.4 x 10(-7) M, respectively. The Bmax for binding (nmol bound/mg protein) was 50.7 and 61.1 for dLO and TMP-OH, respectively, yielding a molar ratio of approximately 1 for both ligands. The ability of dLO and TMP-OH to bind to human-derived alpha 1-acid glycoprotein, rat-derived retinol-binding protein, human protein-1, and bovine beta-lactoglobulin was also studied. These superfamily proteins are generally abundant in plasma, are freely filtered across the glomerulus, and can bind a wide range of ligands. However, neither dLO nor TMP-OH bound to any of the superfamily proteins. In contrast, under identical experimental conditions, alpha 1-acid glycoprotein did bind progesterone (Kd = 10(-6) M), whereas both beta-lactoglobulin and retinol-binding protein bound retinol (Kd = 10(-8) M for both proteins). These results indicate that, under conditions where alpha 2u-globulin superfamily proteins bind to established ligands, the proteins do not interact with hyaline droplet inducing agents. Thus, the interaction between male rat-specific nephrotoxicants and alpha 2u-globulin is unique to this protein. More importantly, these results provide direct evidence that the presence of the alpha 2u-globulin superfamily proteins does not predispose humans to develop hyaline droplet nephropathy and renal cancer from this class of chemicals.
Lehman-McKeeman
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Human & Environmental Safety Division, Miami Valley Laboratories, Procter & Gamble Co., Cincinnati, Ohio 45239.
Caudill
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D000510:Alpha-Globulins / Q000378:metabolism*; D000818:Animals; D000081005:Cyclohexane Monoterpenes; D005260:Female; D006818:Hyalin / Q000187:drug effects*; D007700:Kinetics; D007782:Lactoglobulins / Q000378:metabolism; D008297:Male; D039821:Monoterpenes; D009961:Orosomucoid / Q000378:metabolism; D000439:Pentanols / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding; D051381:Rats; D011916:Rats, Inbred F344; D012177:Retinol-Binding Proteins / Q000378:metabolism; D054839:Retinol-Binding Proteins, Plasma; D013329:Structure-Activity Relationship; D013729:Terpenes / Q000378:metabolism* / Q000494:pharmacology
D000510:Alpha-Globulins; D000081005:Cyclohexane Monoterpenes; D007782:Lactoglobulins; D039821:Monoterpenes; D009961:Orosomucoid; D000439:Pentanols; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma; D013729:Terpenes; C013803:alpha 2u globulin; C028940:limonene-1,2-epoxide; C067352:2,4,4-trimethyl-2-pentanol