Transmembrane domain histidines contribute to regulation of AE2-mediated anion exchange by pH

Author:

Stewart A. K.,Kurschat C. E.,Burns D.,Banger N.,Vaughan-Jones R. D.,Alper S. L.

Abstract

Activity of the AE2/SLC4A2 anion exchanger is modulated acutely by pH, influencing the transporter's role in regulation of intracellular pH (pHi) and epithelial solute transport. In Xenopus oocytes, heterologous AE2-mediated Cl/Cland Cl/HCO3exchange are inhibited by acid pHior extracellular pH (pHo). We have investigated the importance to pH sensitivity of the eight histidine (His) residues within the AE2 COOH-terminal transmembrane domain (TMD). Wild-type mouse AE2-mediated Cl/Clexchange, measured as DIDS-sensitive36Clefflux from Xenopus oocytes, was experimentally altered by varying pHiat constant pHoor varying pHo. Pretreatment of oocytes with the His modifier diethylpyrocarbonate (DEPC) reduced basal36Clefflux at pHo7.4 and acid shifted the pHovs. activity profile of wild-type AE2, suggesting that His residues might be involved in pH sensing. Single His mutants of AE2 were generated and expressed in oocytes. Although mutation of H1029 to Ala severely reduced transport and surface expression, other individual His mutants exhibited wild-type or near-wild-type levels of Cltransport activity with retention of pHosensitivity. In contrast to the effects of DEPC on wild-type AE2, pHosensitivity was significantly alkaline shifted for mutants H1144Y and H1145A and the triple mutants H846/H849/H1145A and H846/H849/H1160A. Although all functional mutants retained sensitivity to pHi, pHisensitivity was enhanced for AE2 H1145A. The simultaneous mutation of five or more His residues, however, greatly decreased basal AE2 activity, consistent with the inhibitory effects of DEPC modification. The results show that multiple TMD His residues contribute to basal AE2 activity and its sensitivity to pHiand pHo.

Publisher

American Physiological Society

Subject

Cell Biology,Physiology

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3