title: Efficient purification of a full length and biochemically active p110Rb, the retinoblastoma gene product.
abstract: Rb protein was purified from recombinant baculovirus-infected Sf9 cells to apparent homogeneity by a simple solubilization and by immunoaffinity column chromatography. It was mainly obtained as p110Rb, the underphosphorylated form of the Rb protein family. This p110Rb was shown to form a specific complex in vitro with SV40 Tag and to bind to double or single stranded DNA. It could also affect Tag helicase activity in a biphasic-dose dependent manner, due to these two biochemical functions. This purification procedure provides sufficient amounts of native Rb protein to further elucidate its role as an anti-oncogene protein and a negative regulator of the cell cycle.
authors.affiliation: Laboratory of Cancer Cell Biology, Nagoya University School of Medicine, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Savoysky
authors.forename: M
authors.initials: M
authors.lastname: Suzuki
authors.forename: K
authors.initials: K
authors.lastname: Tamai
authors.forename: T
authors.initials: T
authors.lastname: Sakurai
authors.forename: T
authors.initials: T
authors.lastname: Ohuchi
authors.forename: T
authors.initials: T
authors.lastname: Akiyama
authors.forename: K
authors.initials: K
authors.lastname: Kojima
authors.forename: S
authors.initials: S
authors.lastname: Yoshida
mesh_terms: D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism; D001483:Base Sequence; D002460:Cell Line; D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007163:Immunosorbent Techniques; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000302:isolation & purification* / Q000378:metabolism; D016622:Silver Staining
chemical_list: D000952:Antigens, Polyomavirus Transforming; D011994:Recombinant Proteins; D016160:Retinoblastoma Protein; D004247:DNA