Protein tyrosine kinase in rat brain: neonatal rat brain expresses two types of pp60c-src and a novel protein tyrosine kinase.
Using angiotensin I as a substrate, the activity of protein tyrosine kinase was determined in various rat tissues, and its developmental change in rat brain was investigated. The specific activity was shown to be the highest in the brain among the tissues examined in neonatal rats, while it was the highest in the spleen in adult rats. In the brain, the activity varied during development and was the highest in the first postnatal week. To identify the protein tyrosine kinase and examine its relationship with pp60c-src, which is known to be highly expressed in neuronal cells, we attempted to characterize the enzyme from neonatal and adult rat brain, using poly(Glu,Tyr) as a substrate. Neonatal brain was found to express two types of pp60c-src and a novel protein tyrosine kinase to almost the same level, while adult brain expressed pp60c-src predominantly. The neonatal type of pp60c-src and the novel enzyme were designated as pp60nc-src and N-PTK in the present study, respectively. pp60c-src, pp60nc-src, and N-PTK were purified about 660-. 370-, and 260-fold from crude homogenate of neonatal brain, respectively, by procedures including sequential column chromatography on DEAE cellulose, hydroxylapatite, Ultrogel AcA44, and poly(Glu,Tyr) Sepharose. N-PTK behaved as a molecule with apparent Mr = 50,000 on Ultrogel AcA44 gel filtration chromatography. It was not immunoprecipitated by anti-pp60c-src antiserum and did not phosphorylate IgG heavy chain of anti-pp60c-src antibody. It required mainly Mn2+ for activity and phosphorylated tyrosine-containing polyamino acids and synthetic peptides such as angiotensin II and RR-SRC peptide.(ABSTRACT TRUNCATED AT 250 WORDS)
D000375:Aging / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism*; D000911:Antibodies, Monoclonal; D001921:Brain / Q000201:enzymology* / Q000254:growth & development; D002413:Cations, Divalent; D002846:Chromatography, Affinity; D007700:Kinetics; D008566:Membranes / Q000032:analysis; D015688:Oncogene Protein pp60(v-src); D010766:Phosphorylation; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000032:analysis* / Q000037:antagonists & inhibitors; D051381:Rats; D011919:Rats, Inbred Strains; D012191:Retroviridae Proteins / Q000032:analysis*; D013379:Substrate Specificity
D000911:Antibodies, Monoclonal; D002413:Cations, Divalent; D012191:Retroviridae Proteins; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src)