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The interactome of the N-terminus of band 3 regulates red blood cell metabolism and storage quality
bioRxiv - Biochemistry Pub Date : 2020-11-30 , DOI: 10.1101/2020.11.30.404756
Aaron Issaian , Ariel Hay , Monika Dzieciatkowska , Domenico Roberti , Silverio Perrotta , Zsuzsanna Darula , Jasmina Redzic , Micheal P. Busch , Grier P. Page , Kirk C. Hansen , Elan Z Eisenmesser , James C Zimring , Angelo D’Alessandro

Band 3 (anion exchanger 1, AE1) is the most abundant membrane protein in red blood cells (RBCs), the most abundant cell in the human body. A compelling model based on indirect evidence posits that, at high oxygen saturation, the N term cytosolic domain of AE1 binds to and inhibits glycolytic enzymes, thus diverting metabolic fluxes to the pentose phosphate pathway to generate reducing equivalents. Dysfunction of this mechanism occurs during RBC aging or storage under blood bank conditions, suggesting a role for AE1 in the regulation of blood storage quality and efficacy of transfusion, a life saving intervention for millions of recipients worldwide. Here we leverage two murine models carrying genetic ablations of AE1 to provide the first direct mechanistic evidence of its role in metabolic regulation and blood storage quality. Observations in mice phenocopied those in a human subject lacking expression of AE1 residues 1 to 11 (band 3 Neapolis), while common polymorphisms in the region coding for AE1 residues 1 to 56 increased susceptibility to osmotic hemolysis in healthy blood donors. Through thermal proteome profiling and cross-linking proteomics, we provide the first comprehensive analysis of the RBC interactome, with a focus on AE1 residues 1 to 56 and validate recombinant AE1 interactions with glyceraldehyde 3phosphate dehydrogenase (GAPDH). Finally, we show that incubation with a cell penetrating AE1 residues 1 to 56 peptide can rescue the metabolic defect in glutathione recycling and boost post transfusion recoveries of stored RBCs from healthy human donors and genetically ablated mice, paving the way for the in vivo metabolic manipulation of RBCs facing oxidant stress, a landmark of many diseases.

中文翻译:

带3 N末端的相互作用基因组调节红细胞的代谢和储存质量

带3(阴离子交换剂1,AE1)是红细胞(RBC)中含量最高的膜蛋白,红细胞是人体中最丰富的细胞。一个基于间接证据的令人信服的模型认为,在高氧饱和度下,AE1的N端胞质结构域会结合并抑制糖酵解酶,从而将代谢通量转移至戊糖磷酸途径,从而产生还原当量。该机制的功能障碍发生在RBC老化或在血库条件下储存期间,这表明AE1在调节血液储存质量和输血功效中起作用,这是全球数百万接收者的救生干预措施。在这里,我们利用携带AE1基因消融的两个鼠模型来提供其在代谢调节和血液存储质量中作用的第一个直接的机械证据。在小鼠的表型中观察到的是在没有表达AE1残基1至11(带3 Neapolis)的人类受试者中的小鼠,而在编码AE1残基1至56的区域中常见的多态性增加了健康献血者对渗透性溶血的敏感性。通过热蛋白质组分析和交联蛋白质组学,我们提供了RBC相互作用组的第一个综合分析,重点是AE1残基1到56,并验证了重组AE1与3磷酸甘油醛脱氢酶(GAPDH)的相互作用。最后,我们发现,与穿透AE1 1至56肽的细胞一起孵育可以挽救谷胱甘肽循环中的代谢缺陷,并提高健康人捐赠者和基因消融小鼠的RBC输血后的回收率,
更新日期:2020-12-01
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