title: Determination of phosphorylated amino acid residues of Rab8 from Bombyx mori.
abstract: The Rab family of small GTPases are key regulators of membrane trafficking. Partially purified Rab8 from Bombyx mori (BRab8) was phosphorylated by protein kinase C in mammalian cells in vitro. To determine which of the seven serines and four threonines are phosphorylated, we generated deletion and site-directed mutants of BRab8, inserted them in Escherichia coli, partially purified the encoded fusion proteins by affinity chromatography, and examined their phosphorylation by protein kinase C in vitro. We found that Ser-132 of BRab8 was specifically phosphorylated by protein kinase C. In addition, Western blotting using an antiserum against BRab8 and in-gel staining for phosphorylated proteins revealed that BRab8 is phosphorylated in vivo.
authors: Laboratory of Biological Chemistry, Department of Biofunctional Chemistry, Faculty of Agriculture, Kobe University, Kobe, Japan. unotom@kobe-u.ac.jp
authors: Tomohide
authors: T
authors: Uno
authors: Takuya
authors: T
authors: Nakada
authors: Sota
authors: S
authors: Okamaoto
authors: Masahiko
authors: M
authors: Nakamura
authors: Mamoru
authors: M
authors: Matsubara
authors: Hiromasa
authors: H
authors: Imaishi
authors: Hiroshi
authors: H
authors: Yamagata
authors: Kengo
authors: K
authors: Kanamaru
authors: Michihiro
authors: M
authors: Takagi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D012831:Bombyx / Q000201:enzymology / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*; D020691:rab GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D012694:Serine; D011493:Protein Kinase C; D020691:rab GTP-Binding Proteins