Detection of a purine analogue-sensitive kinase in frog sciatic nerves--possible involvement in nerve regeneration.
This study reports the existence of a purine analogue-sensitive protein kinase in adult frog sciatic nerves. Cell-free supernatants of homogenized regenerating sciatic nerves were found to contain a phosphoprotein (MW 90 kDa, referred to as PP90), that was phosphorylated to a much higher degree than in normal, uninjured nerves. The spatial and temporal characteristics of PP90 phosphorylation suggested a relationship with the injury-induced proliferation of support cells of the regenerating nerve, i.e. its appearance and increment over time correlated with that of [3H]thymidine incorporation in the nerve. PP90 was phosphorylated under conditions that excluded enzyme activities due to Ca2+/calmodulin kinases, cyclic nucleotide-dependent kinases or protein kinase C. On the other hand, the phosphorylation could be selectively inhibited by the purine analogues adenosine, 2-aminopurine and 6-thioguanine (6-TG). The latter was the most potent and gave complete inhibition at 50 microM. Addition of histone H1 to the cell-free assay stimulated the phosphorylation of several proteins in both normal and regenerating nerves. The stimulation could be blocked by 6-TG, indicating the presence of a purine-sensitive kinase also in uninjured nerves. Separate experiments showed that in vitro regeneration of the frog sciatic sensory axons, as well as the proliferation of the support cells, was inhibited by 100 microM 6-TG.
Svensson
B
B
Department of Animal Physiology, University of Lund, Sweden.
Biessels
G J
GJ
Ekström
A R
AR
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D066298:In Vitro Techniques; D008970:Molecular Weight; D009416:Nerve Regeneration / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000166:cytology; D010766:Phosphorylation; D010770:Phosphotransferases / Q000737:chemistry / Q000378:metabolism*; D011687:Purines / Q000494:pharmacology*; D011896:Rana temporaria; D012584:Sciatic Nerve / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D013866:Thioguanine / Q000494:pharmacology; D013997:Time Factors; D014018:Tissue Distribution
D011687:Purines; D010770:Phosphotransferases; D013866:Thioguanine