Purification and cDNA cloning of ARPP-16, a cAMP-regulated phosphoprotein enriched in basal ganglia, and of a related phosphoprotein, ARPP-19.
ARPP-16 (cAMP-regulated phosphoprotein of Mr = 16,000) is a substrate for cAMP-dependent protein kinase and is enriched in the basal ganglia. ARPP-16 has been purified to homogeneity from the soluble fraction of bovine caudate nuclei. An additional substrate for cAMP-dependent protein kinase of Mr = 19,000 (ARPP-19) was found to cross-react with rabbit anti-serum prepared against purified ARPP-16. Immunological analysis indicated that ARPP-16 was enriched in the basal ganglia while ARPP-19 was present in similar levels in all brain regions studied and was also present in non-neuronal tissues. ARPP-19 was also purified to homogeneity from bovine caudate nucleus cytosol. Using oligonucleotide probes based on the partial amino acid sequence of purified ARPP-16, cDNA clones for ARPP-16 and ARPP-19 were isolated from a bovine caudate nucleus cDNA library and sequenced. The predicted amino acid sequences of the two proteins were identical except that ARPP-19 had an additional 16 amino acids at the NH2-terminal. The two cDNA clones share an identical 3'-untranslated region of 756 nucleotides. The cDNA clone for ARPP-16 contained 806 additional nucleotides located 3' to this common sequence. The 5'-untranslated regions of the two clones were entirely different. The results suggest the possibility that ARPP-16 and ARPP-19 are produced by alternative transcription and splicing.
Horiuchi
A
A
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
Williams
K R
KR
Kurihara
T
T
Nairn
A C
AC
Greengard
P
P
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001479:Basal Ganglia / Q000032:analysis*; D001483:Base Sequence; D001923:Brain Chemistry; D002417:Cattle; D002421:Caudate Nucleus / Q000032:analysis; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000494:pharmacology*; D003600:Cytosol / Q000032:analysis; D004247:DNA / Q000235:genetics; D015151:Immunoblotting; D007163:Immunosorbent Techniques; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000235:genetics / Q000302:isolation & purification*; D011494:Protein Kinases / Q000378:metabolism; D015183:Restriction Mapping; D014018:Tissue Distribution
D000596:Amino Acids; D010750:Phosphoproteins; C064322:cyclic AMP-regulated phosphoprotein 16; C064321:cyclic AMP-regulated phosphoprotein 19; D004247:DNA; D000242:Cyclic AMP; D011494:Protein Kinases