人、牛和大鼠视网膜及松果体中33-kDa蛋白和cDNA的分析。

Analysis of the human, bovine and rat 33-kDa proteins and cDNA in retina and pineal gland.

作者信息

Abe T, Nakabayashi H, Tamada H, Takagi T, Sakuragi S, Yamaki K, Shinohara T

机构信息

Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, NIH, Bethesda, MD 20892.

出版信息

Gene. 1990 Jul 16;91(2):209-15. doi: 10.1016/0378-1119(90)90090-e.

DOI:10.1016/0378-1119(90)90090-e

PMID:2210381

Abstract

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.

摘要

制备了一种针对牛视网膜33 kDa蛋白的单克隆抗体（mAb）。通过mAb筛选以及与cDNA探针杂交，从人、牛和大鼠视网膜以及大鼠松果体的每个cDNA文库中分离出了几个33 kDa蛋白的克隆。测定了四个cDNA序列中的每一个，并从核苷酸序列推导了氨基酸（aa）序列。后两者在大鼠视网膜和大鼠松果体中几乎相同（99.6%），在人、牛和大鼠视网膜中相似（超过87%）。这些cDNA中的每一个都有一个长开放阅读框（ORF），编码245或246个氨基酸。大鼠视网膜和大鼠松果体中推导的氨基酸序列实际上是相同的，人、牛和大鼠视网膜中的序列高度同源（超过88%）。这些蛋白质中每一个的预测相对分子质量（Mr）在人中为28,246，在牛中为28,176，在大鼠视网膜中为28,143，在大鼠松果体中为28,129。每个序列都有一个假定的被A激酶磷酸化的位点；我们已经证实该假定位点是Ser73。这些结果表明，视网膜和松果体中的33 kDa蛋白具有相同的序列和相同的磷酸化位点，并表明该蛋白在视网膜和松果体中的功能作用相同。