Analysis of the human, bovine and rat 33-kDa proteins and cDNA in retina and pineal gland.
A monoclonal antibody (mAb) was produced against a bovine retinal 33-kDa protein. Several clones of 33-kDa protein were isolated from each library of cDNA from human, bovine and rat retinas and rat pineal gland by mAb screening and by hybridization with cDNA probes. Each of the four cDNA sequences was determined and amino acid (aa) sequences were deduced from the nucleotide sequences. The latter were nearly identical in rat retina and rat pineal gland (99.6%) and were similar in human, bovine and rat retina (more than 87%). Each of these cDNAs had one long ORF and encoded 245 or 246 aa. The deduced aa sequences in rat retina and rat pineal gland were virtually identical and the sequences in human, bovine and rat retina were highly homologous (more than 88%). The predicted Mr for each of these proteins was 28,246 in the human, 28,176 in bovine, 28,143 in rat retina, and 28,129 in rat pineal gland. Each of the sequences has a putative site for phosphorylation by A kinase; we have confirmed that the putative site is Ser73. These results show that the 33-kDa proteins in the retina and pineal gland have the same sequences and the same phosphorylation site and suggest that the functional role of this protein is the same in the retina and pineal gland.
Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, NIH, Bethesda, MD 20892.
T
T
Abe
H
H
Nakabayashi
H
H
Tamada
T
T
Takagi
S
S
Sakuragi
K
K
Yamaki
T
T
Shinohara
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002417:Cattle; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D005136:Eye Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010750:Phosphoproteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D010870:Pineal Gland / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D012160:Retina / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction
D000911:Antibodies, Monoclonal; D005136:Eye Proteins; D010750:Phosphoproteins; C065972:retinal protein, 33 kDa; D004247:DNA; D011494:Protein Kinases