Brown C D, Dunk C R, Turnberg L A
Department of Medicine, Hope Hospital, University of Manchester School of Medicine, Salford, United Kingdom.
Am J Physiol. 1989 Oct;257(4 Pt 1):G661-7. doi: 10.1152/ajpgi.1989.257.4.G661.
Transport mechanisms for Cl and HCO3 anions in the apical membrane of rat duodenal enterocytes have been characterized using brush-border membrane vesicles. 36Cl uptake was stimulated by outwardly directed gradients of OH, HCO3, and Cl anions. The anion exchanger was sensitive to inhibition by 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) (Ki, 1.3 mmol/l), 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), and furosemide. The process was electroneutral, since voltage clamping the membrane potential to 0 mV or applying a large inside-positive potential had no effect on the magnitude of uptake. The kinetic properties of the exchanger were measured and an apparent Km of 9.8 mM Cl and a Vmax of 134 nmol.mg protein-1.4 s-1 were found. In addition, an electrogenic conductive component of 36Cl uptake was found. This component was dependent on an inside-positive membrane potential and was inhibited by the Cl channel blocker diphenylamine-2-carboxylate. SITS, DIDS, and furosemide had no effect on the electrogenic component of 36Cl uptake. An apparent anion selectivity of SCN greater than I greater than Br greater than Cl greater than HCO3 greater than SO4 greater than Glu greater than PO4 was found. These results support the presence of both Cl-HCO3 exchange and a conductive anion channel in the apical membrane of rat duodenal enterocytes.
利用刷状缘膜囊泡对大鼠十二指肠肠上皮细胞顶端膜中氯离子(Cl)和碳酸氢根离子(HCO₃)的转运机制进行了表征。外向的氢氧根离子(OH)、碳酸氢根离子和氯离子梯度刺激了³⁶Cl的摄取。阴离子交换体对4-乙酰氨基-4'-异硫氰基芪-2,2'-二磺酸(SITS)(抑制常数Ki为1.3 mmol/L)、4,4'-二异硫氰基芪-2,2'-二磺酸(DIDS)和呋塞米敏感。该过程是电中性的,因为将膜电位钳制在0 mV或施加较大的内正电位对摄取量大小没有影响。测量了交换体的动力学特性,发现其对Cl的表观米氏常数(Km)为9.8 mM,最大反应速度(Vmax)为134 nmol·mg蛋白⁻¹·4 s⁻¹。此外,还发现了³⁶Cl摄取的一个生电传导成分。该成分依赖于内正膜电位,并受到Cl通道阻滞剂二苯胺-2-羧酸盐的抑制。SITS、DIDS和呋塞米对³⁶Cl摄取的生电成分没有影响。发现阴离子选择性顺序为硫氰酸根离子(SCN)大于碘离子(I)大于溴离子(Br)大于氯离子大于碳酸氢根离子大于硫酸根离子(SO₄)大于谷氨酸根离子(Glu)大于磷酸根离子(PO₄)。这些结果支持大鼠十二指肠肠上皮细胞顶端膜中存在Cl-HCO₃交换和一个传导性阴离子通道。