Soluble protein kinase fractions from DEAF-cellulose chromatography

✍ Scribed by Ralph F. Alderson; Paul Y. Sze; Max Costa

• Book ID: 104674480
• Publisher: Springer
• Year: 1982
• Tongue: English
• Weight: 380 KB
• Volume: 49
• Category: Article
• ISSN: 0300-8177
• DOI: 10.1007/bf00231176

✦ Synopsis

The cytosol fraction from rat midbrain was chromatographed on DEAE-cellulose with a linear NaCl gradient (0-0.3 M). Two peaks of protein kinase activity were obtained when assayed with either histone or casein. A similar elution profile of the kinase activity was obtained from rat heart. The first peaks from midbrain and heart were compared in terms of their dependency upon cAMP and sensitivity to the endogenous protein kinase inhibitor. Neither of the two substances had an effect on the activity of the brain kinase. Furthermore, the dissociability of the midbrain and heart enzymes in the presence of cAMP or histone was compared by DEAE-cellulose chromatography. The heart enzyme was dissociated into a catalytic subunit characteristic of a cAMP-dependent protein kinase, whereas the brain kinase was totally unaffected by the cAMP or histone. The results of these tests indicate that although the elution profiles from DEAE-cellulose are similar between midbrain and heart, the first peak from brain contains a protein kinase that appears to be cAMP independent.