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A sphingosine-dependent protein kinase that specifically phosphorylates 14-3-3 (SDK1) is identified as the kinase domain of PKCdelta: a preliminary note
Authors:Hamaguchi Akikazu  Suzuki Erika  Murayama Kimie  Fujimura Tsutomu  Hikita Toshiyuki  Iwabuchi Kazuhisa  Handa Kazuko  Withers Donald A  Masters Shane C  Fu Haian  Hakomori Senitiroh
Institution:Department of Pathobiology, University of Washington, Seattle, WA, USA.
Abstract:A specific protein kinase that phosphorylates Ser60, Ser59, or Ser58 of 14-3-3beta, eta, or zeta, respectively, only in the presence of sphingosine (Sph) or N,N-dimethyl-Sph (DMS), was termed "sphingosine-dependent protein kinase-1" (SDK1) J. Biol. Chem. 273(34) (1998) 21834]. We have now identified SDK1 as a protein having the same amino acid sequence as in the C-terminal-half kinase domain of PKCdelta, with approximately 40 kDa molecular mass, based on large-scale purification of a protein from rat liver, and partial sequence using three different combinations of LC-MS or LC-MS/MS with respective search engine. PKCdelta did not display any SDK1 activity and PKCdelta activity was inhibited by Sph and DMS. However, strong SDK1 activity, only in the presence of Sph or DMS, became detectable when PKCdelta was incubated with caspase-3, which releases the approximately 40 kDa kinase domain.
Keywords:LC-MS/MS  Amino acid sequence  Sphingosine  N  N-Dimethylsphingosine  Sphingosine-dependent protein kinase  14-3-3  Protein kinase Cδ  Caspase-3  Catalytic domain  Phosphorylation at dimer interface
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