Three phosphorylation sites in elongation factor 2

FEBS Letters

Natapov, Pavel G.⁴  Motuz, Lyudmila P.⁴  Averbuch, Lidiya J.⁴  Hardesty, Boyd²  Kramer, Gisela²  Szyszka, Ryszard¹  Ovchinnikov, Lev P.⁴  Wettenhall, Richard E.H.³ 

[1] Department of Molecular Biology, University of Maria Curie Sklodowska, 20-033 Lublin, Poland;Clayton Foundation Biochemical Institute, and Department of Chemistry, The University of Texas at Austin, Austin, TX 78712, USA;The Russell Grimvade School of Biochemistry, The University of Melbourne, Parkville, Victoria 3052, Australia;Institute of Protein Research, Academy of Sciences of the USSR, 142292 Pushchino, Moscow Region, USSR

关键词: Elongation factor 2;    Phosphorylation;    EF-2 kinase;    Tryptic phosphopeptide;    EF-1;    EF-2;    elongation factor 1;    elongation factor 2;    DEAE-cellulose;    diethylaminoethyl cellulose;    TPCK;    L-1-tosyl-amino-2-phenylethyl chloromethyl ketone;    SDS;    sodium dodecyl sulfate;    Hepes;    4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid;    DTT;    dithiothreitol;    TCA;    trichloroacetic acid;    RP-HPLC;    reversed-phase high performance liquid chromatography;    TFA;    trifluoroacetic acid;    EDTA;    ethylenediaminetetraacetic acid;    PTH;    2-phenyl-5-thio-hydantoin;    MES;    2-(N-morpholino) ethanesulfonic acid;    ATZ;    2-anilino-5-thiazolinone;   

DOI  :  10.1016/0014-5793(90)81473-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Elongation factor 2 (EF-2) of rabbit reticulocytes was phosphorylated in vitro by incubation with partially purified EF-2 kinase and (γ³²P)ATP. After exhaustive tryptic hydrolysis 4 phosphopeptides were revealed by two-dimensional peptide mapping. The phosphopeptides were isolated by high performance liquid chromatography and sequenced. A comparison of the primary structure of the phosphopeptides with that of EF-2 showed that all 4 phosphopeptides originated from one region of EF-2 located near the N-terminus that contains 3 threonine residues: Thr-53, Thr-56, Thr-58. A direct estimation of localization of radioactive phosphate in the phosphopeptides demonstrated that all the enumerated threonine residues in EF-2 can be phosphorylated in vitro.

【 授权许可】

Unknown