title: The slowpoke channel binding protein Slob from Drosophila melanogaster exhibits regulatable protein kinase activity.
abstract: The slowpoke channel binding protein Slob from Drosophila melanogaster contains a putative protein kinase domain within its amino acid sequence. We find that Slob exhibits weak and barely detectable protein kinase activity in vitro, as evidenced by autophosphorylation and by phosphorylation of exogenously added histone as substrate. The phosphorylation of histone is enhanced markedly when Slob is pretreated with the catalytic subunit of cyclic AMP-dependent protein kinase (PKAc). Mass spectrometric and mutational analysis demonstrates that the major site of phosphorylation by PKAc within Slob is serine 54. The enhancement of Slob kinase activity by PKAc pretreatment is eliminated when serine 54 in Slob is mutated to alanine (S54A). Furthermore, Slob kinase activity is enhanced in an S54E mutant that mimics phosphorylation at serine 54, and there is no further enhancement of S54E Slob kinase activity by pretreatment with PKAc. The results are consistent with the hypothesis that Slob exhibits regulatable protein kinase activity, whose activity is enhanced by phosphorylation at serine 54.
authors.affiliation: Department of Neuroscience, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
authors.forename: Haoyu
authors.initials: H
authors.lastname: Zeng
authors.forename: Hong
authors.initials: H
authors.lastname: Fei
authors.forename: Irwin B
authors.initials: IB
authors.lastname: Levitan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D004591:Electrophoresis, Polyacrylamide Gel; D006657:Histones / Q000378:metabolism; D013058:Mass Spectrometry; D010766:Phosphorylation; D017354:Point Mutation; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*
chemical_list: D029721:Drosophila Proteins; D006657:Histones; D015221:Potassium Channels; C111538:Slob protein, Drosophila; D011494:Protein Kinases