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首页> 外文期刊>Biochemistry >AE2 anion exchanger polypeptide is a homooligomer in pig gastric membranes: a chemical cross-linking study.
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AE2 anion exchanger polypeptide is a homooligomer in pig gastric membranes: a chemical cross-linking study.

机译：AE2阴离子交换剂多肽是猪胃膜中的均聚物：一种化学交联研究。

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摘要

Although considerable information is available on the oligomeric states of the AE1 (band 3) anion exchanger, little is known about the physiological state of the polypeptides encoded by the nonerythroid AE genes, AE2 and AE3. We have previously characterized the proteolytic susceptibility of native pig gastric AE2. In the course of studies in which pig gastric membranes were treated with the AE2 transport antagonist, DIDS, we noted evidence for cross-linking of AE2 proteolytic fragments to higher-order oligomeric forms. We have characterized the ability of DIDS and of selected N-hydroxysuccinimide cross-linking agents to increase the proportion of SDS-resistant oligomers of pig gastric AE2 and its proteolytic fragments. Cross-linking exhibited time and concentration dependence. N-Terminal protein sequencing proved that DIDS treatment created AE2 homodimers. Putative homotetramers were also observed. Protomers were cross-linked via residues within the C-terminal 40 kDa of AE2. Prior proteolytic cleavage of AE2 in membranes resulted in decreased yield of subsequently cross-linked products. AE2 cross-linking could not be detected in membranes pretreated by hypotonic wash and freeze-thaw. The results are interpreted in light of the deduced amino acid sequence of the transmembrane domain of pig AE2.
机译：尽管可以获得关于AE1（带3）阴离子交换剂的低聚状态的大量信息，但是对于由非红血球AE基因AE2和AE3编码的多肽的生理状态知之甚少。我们先前已经表征了天然猪胃AE2的蛋白水解敏感性。在用AE2转运拮抗剂DIDS处理猪胃膜的研究过程中，我们注意到了AE2蛋白水解片段与高阶寡聚体形式交联的证据。我们已经表征了DIDS和选定的N-羟基琥珀酰亚胺交联剂增加猪胃AE2及其蛋白水解片段的SDS抗性低聚物比例的能力。交联表现出时间和浓度依赖性。 N末端蛋白质测序证明DIDS处理可产生AE2同型二聚体。还观察到推定的同四聚体。启动子通过AE2的C末端40kDa内的残基交联。膜上AE2的事先蛋白水解裂解导致后续交联产物的收率降低。在低渗洗涤和冻融预处理的膜中未检测到AE2交联。根据猪AE2跨膜结构域的推导氨基酸序列来解释结果。

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