title: Phospho-regulation of human protein kinase Aurora-A: analysis using anti-phospho-Thr288 monoclonal antibodies.
abstract: Mammalian Aurora-A is related to a serine/threonine protein kinase that was originally identified by its close homology with Saccharomyces cerevisiae Ipl1p and Drosophila melanogaster aurora that are key regulators in the orchestration of mitotic events. The protein level of Aurora-A, its peak kinase activity during mitosis, and its activation have been attributed to phosphorylation. Here we show that this enzyme is an arginine-directed kinase and define its substrate specificity. We also found that Thr288 within the activation loop is a critical residue for activating phosphorylation events in vitro and that it is spatiotemporally restricted to a brief window at mitosis on duplicated centrosomes and on spindle microtubules proximal to the poles in vivo. Immunodepletion assays indicated that an upstream kinase(s) of Aurora-A might exist in mammalian cells in addition to autophosphorylation. Furthermore, human activated Aurora-A forms complexes with the negative regulator protein serine/threonine phosphatase type 1 (PP1) that was negatively phosphorylated on Thr320. Interestingly, phospho-specific Aurora-A monoclonal antibodies restrain Aurora-A kinase activity in vitro, providing further therapeutic avenues to explore.
authors.affiliation: Department of Biochemistry II, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Ohashi
authors.forename: G
authors.initials: G
authors.lastname: Sakashita
authors.forename: R
authors.initials: R
authors.lastname: Ban
authors.forename: M
authors.initials: M
authors.lastname: Nagasawa
authors.forename: H
authors.initials: H
authors.lastname: Matsuzaki
authors.forename: Y
authors.initials: Y
authors.lastname: Murata
authors.forename: H
authors.initials: H
authors.lastname: Taniguchi
authors.forename: H
authors.initials: H
authors.lastname: Shima
authors.forename: K
authors.initials: K
authors.lastname: Furukawa
authors.forename: T
authors.initials: T
authors.lastname: Urano
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D064127:Aurora Kinases; D006801:Humans; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D000911:Antibodies, Monoclonal; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases