| First Author | Masucci MT | Year | 1989 |
| Journal | Tumori | Volume | 75 |
| Issue | 3 | Pages | 202-10 |
| PubMed ID | 2773071 | Mgi Jnum | J:27968 |
| Mgi Id | MGI:75602 | Doi | 10.1177/030089168907500303 |
| Citation | Masucci MT, et al. (1989) Purification to homogeneity of the major 4S PAH binding protein from non responsive DBA/2N mouse liver by affinity chromatography. Tumori 75(3):202-10 |
| abstractText | DBA/2N is a genetically non responsive inbred strain of mice in which administration of polycyclic aromatic hydrocarbons (PAHs) does not induce microsomal monooxygenase activity. DBA/2N mouse liver cytosol contains a polycyclic aromatic hydrocarbon-binding protein that sediments, in a sucrose gradient, at 4S (4S PAH-BP). Its binding kinetic and physicochemical properties indicate that this protein is practically indistinguishable from the 4S PAH-BP identified and characterized in liver cytosol of rats and other PAH responsive rodents including C57 B1/6J mice. 4S PAH-BP was purified to homogeneity from DBA/2N mouse liver by ammonium sulfate fractionation of the cytosol, followed by Sephadex G-200 chromatography and, finally, affinity chromatography using 1-aminopyrene-Sepharose 6B. This procedure yielded about 50 micrograms of protein from 50-60 g of mouse liver, with a recovery of 18%. 4S PAH-BP as a complex with 3H-(benzo-a-pyrene) was more than 99% pure. A single band was seen on polyacrylamide gel electrophoresis under non denaturing conditions. H-BaP comigrated with the protein band. 3H-BaP bound to the protein was displaced by PAHs with a specificity identical to that obtained using crude cytosol. On electrophoresis in SDS gels, the purified protein migrated as a single protein band with an apparent molecular weight of 40,000. |
