Phosphorylation of threonine residues on cloned fragments of the Dictyostelium myosin heavy chain

FEBS Letters

Scheel, Jochen¹  Noegel, Angelika¹  Wagle, Gertrud¹  Gerisch, Günther¹ 

[1] Max-Planck-Institut für Biochemie, D-8033 Martinsried, FRG

关键词: Myosin phosphorylation;    Protein kinase;    Cell motility;    Cloned myosin tail fragment;    (Dictyostelium discoideum);    CHAPS;    3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate;    DTT;    dithiothreitol;    MHC;    myosin heavy chain;    PMSF;    phenylmethylsulfonyl fluoride;   

DOI  :  10.1016/0014-5793(88)81416-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A tail fragment of Dictyostelium discoideum myosin has been cloned and expressed as a fusion protein with the N-terminal region of MS-2 polymerase. The cloned fragment was phosphorylated with myosin heavy chain kinase II from aggregation-competent D.discoideum cells that specifically phosphorylate threonine residues on the myosin tail. Phosphopeptide maps showed the same site specificity of phosphorylation with the fusion protein as a substrate as with native myosin. An improved assay for the kinase was developed in which the fusion protein is precipitated with a monoclonal antibody that inhibits polymerization of the myosin tails without preventing their phosphorylation. Sites of phosphorylation were tentatively localized to a sequence in the C-terminal region of the heavy chain where four threonine residues are found.

【 授权许可】

Unknown