Partial purification of protein kinase C isoenzymes from rat liver.
Due to the growing number of recently cloned isoenzymes, purification and assay of protein kinase C (PKC) have become increasingly cumbersome. This paper reports the development of a shortened protocol for partial purification and assay of alpha, beta, delta and zeta PKC from rat liver, allowing the determination of a PKC subspecies activity pattern on a single tissue preparation. Calcium-dependent alpha and beta PKC subspecies were resolved by application of a DEAE eluate to a hydroxylapatite column, delta PKC was separated with SP-Sepharose and phenyl-Sepharose chromatography, whereas three column passages were necessary to isolate zeta PKC: DEAE-Sepharose, phenyl-Sepharose and heparin-Sepharose. This procedure allows reproducible separation and assay as well as constant recovery of the four liver PKC isoenzymes.
Perletti
G P
GP
Università degli Studi di Milano, Istituto di Farmacologia, Italy.
D000818:Animals; D002847:Chromatography, Agarose; D007527:Isoenzymes / Q000302:isolation & purification*; D008099:Liver / Q000201:enzymology*; D008297:Male; D011493:Protein Kinase C / Q000302:isolation & purification*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results
D007527:Isoenzymes; D011493:Protein Kinase C