Different regulation by pHi and osmolarity of the rabbit ileum brush-border and parietal cell basolateral anion exchanger.
1. The purpose of this study was to look for evidence of a pH-sensitive modifier site on the parietal cell basolateral anion exchanger, determine the pH range in which allosteric regulation takes place, investigate the effect of the osmolarity on internal pH (pHi) dependence and compare it with that of the ileum brush-border anion exchanger. 2. When the pHi in parietal cell basolateral membrane (BLM) vesicles was increased, the rate of Cl(-)-gradient-driven 36Cl- uptake increased from 6.03 +/- 2.24 to 38.09 +/- 3.33 nmol (mg protein)-1 with the steep increase in anion exchange rates occurring within a narrow pH range between pHi 7.0 and 7.5. This was due to allosteric activation by internal OH- and not due to a change in driving force, since the driving force for maximal exchange rates was provided by the outwardly directed Cl- gradient. 3. The pHi dependency curve of parietal cell BLM anion exchange rates was shifted to the left by 0.25 pH units by increasing the osmolarity of the intra- and extravesicular solutions from 300 to 380 mosmol l-1. Thus cell shrinking may activate the parietal cell anion exchanger without a change in pHi and without phosphorylation of the anion exchanger protein. 4. In ileum brush-border membranes, the pHi-dependent increase in the rate of Cl(-)-gradient-driven 36Cl- uptake was more gradual and the half-maximal anion exchange rate was attained at lower pHi (pH 6.5). Increasing the osmolarity from 300 to 500 mosmol l-1 had no effect on pH dependence. 5. We conclude that the parietal cell basolateral and ileum brush-border anion exchangers possess an internal modifier site for allosteric activation by OH-, but the pH range in which allosteric regulation occurs differs between the two exchangers, as does the effect of an increase in osmolarity. Since current evidence suggests that both the parietal cell basolateral and the ileum brush-border anion exchanger are encoded by the AE2 gene, the differences in pHi dependence between the two may be due to alternative splicing, post-transcriptional modification, or the different membrane environment. 6. The pHi range for allosteric activation found in this study would suggest that for both the ileum and the parietal cell anion exchanger, but especially for the latter, a potentiating effect of the allosteric activation and the HCO3- availability occurs within the physiological pHi range and can cause dramatic increases in maximal anion exchange rates with increasing pHi.
Nader
M
M
II Medizinische Klinik, Technischen Universität München, Germany.
Lamprecht
G
G
Classen
M
M
Seidler
U
U
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D000818:Animals; D017920:Antiporters / Q000378:metabolism*; D001692:Biological Transport; D027963:Chloride-Bicarbonate Antiporters; D006851:Hydrochloric Acid / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007082:Ileum / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008871:Microvilli / Q000378:metabolism*; D009994:Osmolar Concentration; D010295:Parietal Cells, Gastric / Q000378:metabolism*; D011817:Rabbits; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D013997:Time Factors
D017920:Antiporters; D027963:Chloride-Bicarbonate Antiporters; D017923:Sodium-Hydrogen Exchangers; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D006851:Hydrochloric Acid