Molecular and Biochemical Studies on Compensatory Mechanisms for Total Band 3 Deficiency in Japanese Black Cattle

日本黑牛总带 3 缺陷补偿机制的分子和生化研究

基本信息

批准号：
09460145
负责人：
INABA Mutsumi
金额：
$ 7.87万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460145/
关键词：
Red cells Membrane proteins Band 3 Hereditary diseases Anion transport Acid-base balance Membrane skeleton Japanese black cattle

项目摘要

Band 3 has been believed to be essential to survival of mammals. The aim of this research project is to define compensatory mechanisms for total band 3 deficiency in cattle.1) Linkage analyses showed that the genotype for R664X mutation determined by PCR-RFLP coinherited with the red cell phenotype of dominantly inherited HS and band 3 deficiency, demonstrating that R664X mutation is the principal molecular cause for dominant hereditary band 3 deficiency in cattle associated with HS.2) Extensive studies on the red cell membrane proteins demonstrated that the major proximal causes for the membrane instability of homozygous and heterozygous cells appear to be the loss of band 3-ankyrin-spectrin association, and the reduction of spectrin, respectively. Quantitation of mutant mRNA co-injection of normal and mutant RNA into Xenopus oocytes, and in vitro synthesis/immunoprecipitation of normal and the mutant bend 3 demonstrated a dominant-negative effect of the mutant protein in vivo on the expression of normal band 3. A hypothetical possibility for pathogenesis of HS in the affected animals involves : (1) Band 3-independent assembly of membrane skeleton to the plasma membrane. (2) Translocation of reduced normal band 3-ankyrin and their association with spectrin to strengthen interactions between the lipid bilayer and the skeleton in heterozygous but not m homozygous red cells.3) Bovine red cells with total band 3 deficiency possessed anion transport activity mediated by AE2, with substrate specificity an sensitivity to stilbene disulfonate which were extremely lower than those in normal cells. T e rapid anion exchange was not compensated at all, indicating that the function of band 3 is not obligatory to 0ィイD22ィエD2/C0ィイD22ィエD2 exchange.

据信，3条对哺乳动物的生存至关重要。 The aim of this research project is to define compensatory mechanisms for total band 3 deficiency in cattle.1) Linkage analyses showed that the genotype for R664X mutation determined by PCR-RFLP coinherited with the red cell phenotype of dominant inherited HS and band 3 deficiency, demonstrating that R664X mutation is the principal molecular cause for dominant hereditary band 3 deficiency in cattle Associated with HS.2) Extensive studies on红细胞膜蛋白表明，纯合细胞和杂合细胞的膜不稳定性的主要代理似乎分别是3-烷基光谱蛋白缔合的丧失，分别是谱蛋白的减少。将正常和突变RNA突变mRNA共注射到爪蟾卵母细胞中，体外合成/对正常弯曲和突变体弯曲3的体外合成/免疫沉淀表现出了突变蛋白在体内对正常带3表达的显性阴性效应。质膜。 (2) Translocation of reduced normal band 3-ankyrin and their association with spectrin to strengthen interactions between the lipid bilayer and the skeleton in heterozygous but not m homozygous red cells.3) Bovine red cells with total band 3 Deficiency assumed anion transport activity mediated by AE2, with substrate specificity an sensitivity to stilbene disulfonate which were extremely lower than those in normal cells.快速阴离子交换根本没有得到补偿，表明频段3的功能不可与0II D22/C0II D22交换。