期刊
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
卷 1768, 期 5, 页码 1023-1035出版社
ELSEVIER
DOI: 10.1016/j.bbamem.2007.01.003
关键词
cell membrane; hydrogen-ion concentration; sodium; exocytosis; endocytosis
资金
- NIDDK NIH HHS [R01 DK42457, R01 DK042457, F32 DK61178, R01 DK054940, F32 DK061178] Funding Source: Medline
We tested whether NHE3 and NHE2 Na+/H+ exchanger isoforms were recruited to the plasma membrane (PM) in response to changes in ion homeostasis. NHE2-CFP or NHE3-CFP fusion proteins were functional Na+/H+ exchangers when transiently expressed in NHE-deficient PS120 fibroblasts. Confocal morphometry of cells whose PM was labeled with FM4-64 measured the fractional amount of fusion protein at the cell surface. In resting cells, 10-20% of CFP fluorescence was at PM and stable over time. A protocol commonly used to activate the Na+/H+ exchange function (NH4-prepulse acid load sustained in Na+-free medium), increased PM percentages of PM NHE3-CFP and NHE2-CFP. Separation of cellular acidification from Na+ removal revealed that only NHE3-CFP translocated when medium Na+ was removed, and only NHE2-CFP translocated when the cell was acidified. NHE2[NHE3 chimeric proteins demonstrate that the Na+-removal response element resides predominantly in the NHE3 cytoplasmic tail and is distinct from the acidification response sequence of NHE2. (c) 2007 Elsevier B.V. All rights reserved.
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