Protein phosphorylation in Rhodnius prolixus oocytes: identification of a type II casein kinase.
A protein kinase activity in chorionated oocytes of Rhodnius prolixus phosphorylates in vitro vitellin (VT), the major yolk protein. Phosphatase inhibitors including NaF, sodium vanadate, beta-glycerophosphate and okadaic acid did not alter the protein phosphorylation profile to a visible extent. Among the exogenous protein substrates tested, casein was readily phosphorylated, but histones were not. Several different protein kinase activators, including cAMP, Ca2+ plus calmodulin, Ca2+ plus diolein and phosphatidylserine, were added to the reaction media but spermidine was the only effective one, inducing a 2-fold increase in the phosphorylation of VT. A strong inhibition was obtained with nanomolar levels of heparin. The enzyme could also accept GTP as the phosphate donor instead of ATP. These properties identify the major protein kinase activity as a type II casein kinase (CK II). The pH dependence and the effects of mono- and divalent cations on VT phosphorylation were also studied. Gel filtration revealed only one peak of protein kinase activity, with a molecular mass of 170 K, similar to values previously reported in the literature for CK IIs from other organisms.
Silva-Neto
M A
MA
Departmento de Bioquímica Médica, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Oliveira
P L
PL
D000818:Animals; D047388:Casein Kinases; D002364:Caseins / Q000378:metabolism; D002414:Cations, Monovalent / Q000494:pharmacology; D002850:Chromatography, Gel; D004527:Egg Proteins / Q000378:metabolism; D005260:Female; D006160:Guanosine Triphosphate / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D008970:Molecular Weight; D009865:Oocytes / Q000201:enzymology*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism; D012239:Rhodnius / Q000201:enzymology*; D013095:Spermidine / Q000494:pharmacology
D002364:Caseins; D002414:Cations, Monovalent; D004527:Egg Proteins; D006657:Histones; D006160:Guanosine Triphosphate; D006493:Heparin; D011494:Protein Kinases; D047388:Casein Kinases; D010749:Phosphoprotein Phosphatases; D013095:Spermidine