权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Molecular cloning,expression,and chromosomal localization of a human tubulointerstitial nephritis antigen

人肾小管间质性肾炎抗原的分子克隆、表达及染色体定位

基本信息

批准号：
11470177
负责人：
YOSHIOKA Kazuo
金额：
$ 6.4万
依托单位：
Kinki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470177/
关键词：
tibilointerstitial nephritis cDNA follistatin cathepsin B chromosome 6 若年性ネフロン癆

项目摘要

Tubulointerstitial nephritis antigen (TIN-ag) is an extracellular matrix base ment protein which was originally identified as a target antigen involved in anti-tubular basement membrane (TBM) antibody-mediated interstitial nephritis(TIN). Further investigations elucidated that TIN-ag plays a role in renal tubulogenesis and that TIN-ag is defected in hereditary tubulointerstitial disorder such as juvenile nephronophthisis. We previously isolated and characterized 54 dKa glycoprotein as TIN-ag. cDNA encoding rabbit and mouse TIN-ag has recently been identified. In the present study, the cDNA of the human honologue of TIN-ag was cloned and its nucleotid sequence was determined (accession No.AB022277, the DDBJ nucleotide sequence database). Deduced amino acid sequence (476aa) exhibited the presence of a signal peptide (1-18aa), cysteine residues termed follistatinmodule, six potential glycosylation sites, and an ATP/GTP-binding site. Homology search revealed 〜 85% homology with both rabbit and mouse TIN-ag, and also some (〜40%) similarity with C. elegans. Human TIN-ag contained a sequence similar to several classes of extracelular matrix molecules in amino terminal region and to cathepsin family of cysteine proteinases in the carboxyl terminal region. Northern blot analysis revealed exclusive expression of this molecule in human adult and fetal kidney tissues. Using a monoclonal antibody recognizing human TIN-ag, protein expression (〜50 kDa) was identified in cultured COS-1 cells transfected with human TIN-ag cDNA. The human TIN-ag was mapped to chromosome 6p11. 2-12 by fluorescence in situ hybridization. These results may privide further evidence for understanding TIN-ag molecule and clues for gene analysis of juvenile nephronophthisis.

肾小管间质性肾炎抗原（TIN-ag）是一种细胞外基质基底蛋白，最初被鉴定为参与抗肾小管基底膜（TBM）抗体介导的间质性肾炎（TIN）的靶抗原。进一步的研究表明，TIN-ag 在肾小管发生中发挥作用，并且 TIN-ag 在遗传性肾小管间质疾病（如青少年肾结核）中存在缺陷。我们之前分离并表征了 54 dKa 糖蛋白作为 TIN-ag。最近已鉴定出编码兔和小鼠 TIN-ag 的 cDNA。在本研究中，克隆了TIN-ag的人类同源物的cDNA并测定了其核苷酸序列（登录号AB022277，DDBJ核苷酸序列数据库）。推导的氨基酸序列 (476aa) 显示存在信号肽 (1-18aa)、称为卵泡抑素模块的半胱氨酸残基、六个潜在的糖基化位点和 ATP/GTP 结合位点。同源性搜索显示与兔和小鼠 TIN-ag 具有约 85% 的同源性，与秀丽隐杆线虫也有一些 (约 40%) 的相似性。人TIN-ag在氨基末端区域含有与几类细胞外基质分子相似的序列，在羧基末端区域含有与半胱氨酸蛋白酶的组织蛋白酶家族相似的序列。 Northern印迹分析揭示了该分子在成人和胎儿肾组织中的独特表达。使用识别人 TIN-ag 的单克隆抗体，在转染人 TIN-ag cDNA 的培养 COS-1 细胞中鉴定出蛋白质表达 (~50 kDa)。人类 TIN-ag 被定位到染色体 6p11。 2-12 通过荧光原位杂交。这些结果可能为理解TIN-ag分子提供进一步的证据，并为青少年肾结核的基因分析提供线索。