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Isolation and characterization of des(Ala-Lys)calmodulin in porcine brain
Authors:Toshiaki Isobe  Noriaki Ishioka  Tsuneo Okuyama
Institution:Department of Chemistry, Faculty of Science, Tokyo Metropolitan University, Stagaya, Tokyo 158, Japan
Abstract:A calmodulin-like protein -des(Ala-Lys)calmodulin- was isolated from porcine brain extract, and was characterized in comparison to porcine brain calmodulin. Des(Ala-Lys)calmodulin was distinguishable from calmodulin by its slightly faster mobility in 10% polyacrylamide gels without sodium dodecyl sulfate. The protein gave an amino acid composition very similar to calmodulin, and contained one ?-N-trimethyllysyl residue. Comparative peptide mapping of calmodulin and des(Ala-Lys)calmodulin by high performance anion-exchange liquid chromatography, and the subsequent analyses of the isolated peptides, have indicated that des(Ala-Lys)calmodulin lacks the Ala(147)-Lys (148) sequence at the C-terminus of calmodulin. The content of des(Ala-Lys)-calmodulin was about one-tenth of calmodulin.
Keywords:SDS  sodium dodecyl sulfate  EDTA  ethylenediamine-tetraacetic acid  TPCK  L-(1-tosylamido-2-phenyl)ethyl chloromethyl ketone
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