title: Inhibitory effect of H-7, H-8 and polymyxin B on liver protein kinase C-induced phosphorylation of endogenous substrates.
abstract: Inhibitory actions of 1-(5-isoquinolinyl-sulfonyl)-2-methylpiperazine (H-7), N-[2-(methylamine)ethyl]-5-isoquinolinesulfonamide [H-8] and polymyxin B on the calcium-activated, phospholipid-dependent protein kinase (protein kinase C) of rat liver were compared. Using a partially purified liver protein kinase C and an exogenous substrate histone-III S, polymyxin B showed maximum inhibition (IC50, 9.5 microM) followed by H-7 (IC50, 25 microM) and H-8 (IC50, 36 microM). These inhibitors also inhibited protein kinase C-induced phosphorylation of endogenous cytosolic and particulate proteins in a dose-dependent manner though polymyxin B was relatively less effective with the particulate fraction. With the aid of protein kinase-C activators and these inhibitors, seven proteins in cytosolic (Mr 170K, 150K, 43K, 34K, 30K, 25K and 19K daltons) and six proteins in particulate (Mr 150K, 43K, 34K, 25K, 19K and 16K daltons) fractions were identified as probable substrates for protein kinase C in liver. The identity of these proteins remains to be determined.
authors.affiliation: Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.
authors.forename: S
authors.initials: S
authors.lastname: Pugazhenthi
authors.forename: S V
authors.initials: SV
authors.lastname: Mantha
authors.forename: R L
authors.initials: RL
authors.lastname: Khandelwal
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006657:Histones / Q000378:metabolism; D007546:Isoquinolines / Q000494:pharmacology*; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D010718:Phosphatidylserines / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology*; D011112:Polymyxin B / Q000494:pharmacology*; D011113:Polymyxins / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D004075:Diglycerides; D006657:Histones; D007546:Isoquinolines; D010718:Phosphatidylserines; D010750:Phosphoproteins; D010879:Piperazines; D011113:Polymyxins; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C053177:N-(2-(methylamino)ethyl)-5-isoquinolinesulfonamide; D011493:Protein Kinase C; D011112:Polymyxin B; D002118:Calcium; C013965:diolein