Absence of 2'-deoxy-GTP in adult brain tubulin.
Nerve growth factor can stimulate incorporation of 2'-deoxy-GTP into the non-exchangeable nucleotide sites in tubulin and cytoskeletal microtubules of PC12 pheochromocytoma cells and embryonic chick dorsal root ganglion neurons [J. M. Angelastro and D. L. Purich (1992) J. Biol. Chem. 267, 25685-25689]. We replaced and hydrolyzed exchangeable-site GTP and GDP in adult bovine brain tubulin by incubation with the non-hydrolyzable nucleotide analogue 5'-guanylyl-methylenediphosphonate and alkaline phosphatase, thereby allowing us to analyze the non-exchangeable guanine nucleotides for GTP and dGTP. HPLC analysis reveals no evidence of dGTP in adult tubulin, suggesting further that the appearance of dGTP in tubulin and microtubules may be a characteristic of recently dividing neurons in response to nerve growth factor.
Yamauchi
P S
PS
Department of Biochemistry & Molecular Biology, University of Florida College of Medicine, Gainesville 32610-0245.
Purich
D L
DL
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003848:Deoxyguanine Nucleotides / Q000032:analysis*; D014404:Tubulin / Q000737:chemistry*
D003848:Deoxyguanine Nucleotides; D014404:Tubulin; C029603:deoxyguanosine triphosphate