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Purification of a phosphoprotein from rat brain closely related to the 80 kDa substrate of protein kinase C identified in Swiss 3T3 fibroblasts
Author(s) -
Morris Clive,
Rozengurt Enrique
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80840-8
Subject(s) - phosphoprotein , sephadex , protein kinase a , chromatography , size exclusion chromatography , biochemistry , gel electrophoresis , polyacrylamide gel electrophoresis , chemistry , protease , substrate (aquarium) , alanine , microbiology and biotechnology , kinase , amino acid , biology , phosphorylation , enzyme , ecology
A phosphoprotein expressed in rat brain is closely related to the 80 kDa substrate of protein kinase C present in 3T3 cells. The protein kinase C substrates from both sources migrate identically on two‐dimensional gel electrophoresis and give similar phosphopeptide fragments when digested with protease. Using a series of chromatographic steps, including DEAE‐cellulose chromatography, Sephadex G150 gel filtration and reverse phase fast protein liquid chromatography, this phosphoprotein was purified 3800‐fold from rat brain. The preparation appears homogenous by one‐ and two‐dimensional gel electrophoresis, is an effective substrate of protein kinase C and contains a high proportion of the acidic amino acids glutamate and aspartate, and of alanine.