Function and gene analysis of an endoplasmic reticlum-resident Ca^<2+>-binding proptein with multiple EF-hand motifs

具有多个EF-hand基序的内质网驻留Ca^2-结合蛋白的功能和基因分析

• 批准号: 05454623
• 负责人: OZAWA Masayuki
• 金额: $ 3.46万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454623/
• 关键词: Calcium-binding protein; EF-hand; Endoplasmic reticulum; Gene

Reticulocalbin, an endoplasmic reticulum (ER) resident Ca^<2+>-binding protein, is a member of the EF-hand Ca^<2+>-binding protein superfamily [Ozawa, M.and Muramatsu, T.(1993) J.Biol.Chem.268,699-705]. The genomic structure of this unique Ca^<2+>-binding protein was analyzed. Southern blot analysis of mouse genomic DNA revealed that there is a single copy of the reticulocalbin gene per hapliod genome. The gene spans over 13 kilobase pairs and encodes six separate exons. Thus, reticulocalbin differs from the cytosolic Ca^<2+>-binding protein calbindin D28 which also has six EF-hand motif domains but the gene for which is divided into 11 exons. Comparison of the gene organization of reticulocalbin with that of other EF-hand proteins revealed that reticulocalbin diverged very early from other members of the EF-hand protein superfamily. Consistent with the observation that reticulocalbin is expressed by different types of cells, the 5'-flanking region of the gene contains a GC-rich (more … More than 80%) area including two Spl binding motifs and a CCAAT box, but no apparent TATA box was present between the CCAAT box and the transcription start site, a feature being found in many housekeeping genes.We have cloned the human homologue of reticulocalbin. The sequence of this clone revealed a novel protein with 95 percent identity in ammino acid sequence to the mouse reticulocalbin, indicating that this molecule has been evolutionarily conserved in mammals. As was found for the mouse reticulocalbin, the human homologue showed the six repeats of a domain containing EF-hand motifs. Interestingly, the conservation of amino acid sequence was not restricted to the Ca^<2+>-binding motifs, consistent with the possibility that reticulocalbin plays some role (s) besides Ca^<2+>-binding. As was found for the mouse homologue, the protein has the HDEL sequence at its carboxy terminus instead of the KDEL sequence, which is more common as a signal to retain resident proteins in the ER of animal cells. The conservation of the HDEL sequence in reticulocalbin in both species raises the possibility that this sequence has some roles in the function (s) of this protein family. Less

网织红蛋白是一种内质网（ER）Ca^<2+>结合蛋白，是EF-手Ca^<2+>结合蛋白超家族的成员[Ozawa，M.and Muramatsu，T.（1993）J.Biol.Chem.268，699 -705]。对这种独特的Ca^<2+>结合蛋白的基因组结构进行了分析。小鼠基因组DNA的Southern印迹分析显示，每个单倍体基因组中存在单拷贝的网织红蛋白基因。该基因跨越超过13个内切酶对，并编码6个独立的外显子。因此，网织红蛋白与胞质Ca^2+结合蛋白钙结合蛋白D28不同，后者也有6个EF-手形基序结构域，但其基因分为11个外显子。网织红蛋白与其他EF-手蛋白的基因组织的比较显示，网织红蛋白从EF-手蛋白超家族的其他成员分化很早。与网织白蛋白由不同类型的细胞表达的观察结果一致，该基因的5 '侧翼区含有富含GC的（更多） ...更多信息 80%以上的区域包含两个Spl结合基序和一个CCAAT盒，但在CCAAT盒和转录起始位点之间没有明显的TATA盒，这是许多管家基因的特征。该克隆的序列揭示了一种新的蛋白质，其氨基酸序列与小鼠网织红蛋白具有95%的同一性，表明该分子在哺乳动物中进化保守。正如在小鼠网织红蛋白中发现的那样，人类同源物显示出含有EF-手状基序的结构域的六个重复。有趣的是，氨基酸序列的保守性并不局限于Ca^2+结合基序，这与网织红蛋白除了Ca^2+结合之外还发挥某些作用的可能性相一致。正如在小鼠同源物中发现的，该蛋白质在其羧基末端具有HDEL序列而不是KDEL序列，其更常见地作为将驻留蛋白质保留在动物细胞ER中的信号。网织红蛋白中HDEL序列在两个物种中的保守性提高了该序列在该蛋白质家族的功能中具有某些作用的可能性。少

项目成果

Ozawa,M,and Muramatsu,T.: "Reticulocalbin,a novel endoplasmic reticulum resident Ca^<2+>- binding protein with multiple EF hand motifs and a carboxy1 terminal HDEL sequence" J.Biol.Chem.268. 699-705 (1993)

Ozawa,M 和 Muramatsu,T.：“Reticulocalbin，一种新型内质网驻留 Ca^2-结合蛋白，具有多个 EF 手基序和羧基 1 末端 HDEL 序列”J.Biol.Chem.268。

小澤政之: "新しいカルシウム結合蛋白質:レティキュロカルビン(Reticulocalbin)" 生体の科学 特集-カルシウムイオンを介した調節機構の新しい問題点. 44. 224-225 (1993)

Masayuki Ozawa：“新钙结合蛋白：Reticulocalbin”生物科学专题 - 钙离子介导的调节机制的新问题 44. 224-225 (1993)

小澤政之: "レティキュロカルビン(Reticulocalbin)" CLINICAL CALCIUM 特集:細胞の機能制御とカルシウム22GD03:3. 1504-1505 (1993)

Masayuki Ozawa：“Reticulocalbin”临床钙专题：细胞功能控制和钙 22GD03:3 (1993)。

Ozawa,M.: "Cloning of a human homologue of mouse reticulocalbin reveals conservation of structural domains in the novel endoplasmic reticulum-resident Ca^<2+>-binding protein with multiple EF-hand motifs" J.Biochem.,. in press. (1995)

Ozawa,M.：“小鼠 reticulocalbin 的人类同源物的克隆揭示了具有多个 EF-hand 基序的新型内质网驻留 Ca^2 结合蛋白中结构域的保守性”J.Biochem.,。

Ozawa, M.and Muramatsu, T.: "Reticulocalbin, a novel endoplasmic reticulum resident Ca^<2+>-binding protein with multiple EF hand motifs and a carboxyl terminal HDEL sequence" J.Biol.Chem.268. 699-705 (1993)

Ozawa，M.和Muramatsu，T.：“Reticulocalbin，一种新型内质网驻留Ca 2+ 结合蛋白，具有多个EF手基序和羧基末端HDEL序列”J.Biol.Chem.268。