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兔回肠刷状缘膜囊泡的pH梯度刺激硫酸盐转运:SO4-OH交换的证据

pH gradient-stimulated sulfate transport by rabbit ileal brush-border membrane vesicles: evidence for SO4-OH exchange.

作者信息

Schron C M, Knickelbein R G, Aronson P S, Della Puca J, Dobbins J W

出版信息

Am J Physiol. 1985 Nov;249(5 Pt 1):G607-13. doi: 10.1152/ajpgi.1985.249.5.G607.

DOI:10.1152/ajpgi.1985.249.5.G607
PMID:4061648
Abstract

In the presence of a pH gradient (7.7 inside, 5.5 outside), the initial velocity of SO4 uptake by rabbit ileal brush-border membrane (BBM) vesicles was markedly stimulated compared with uptake in the absence of a pH gradient. Under pH gradient conditions, SO4 was transiently accumulated at a concentration 13-fold higher than at equilibrium ("overshoot"). Superimposition of a HCO3 gradient did not further stimulate the initial velocity of SO4 uptake compared with a pH gradient alone. Evidence that this pH gradient-stimulated SO4 uptake represented SO4-OH exchange included lack of sensitivity of SO4 transport to alterations of the membrane potential; 85-95% inhibition of SO4 uptake by the anion exchange inhibitors DIDS and SITS; and saturation kinetics (Km for SO4 = 0.475 +/- 0.054 mM; Vmax = 4.1 +/- 0.1 nmol SO4 X mg prot-1 X min-1). Sulfate did not inhibit pH gradient-stimulated 36Cl uptake, indicating that SO4-OH and Cl-HCO3(OH) are different exchangers. When BBM vesicles were compared with basolateral membrane (BLM) vesicles, pH gradient-stimulated SO4 uptake was found predominantly in the BBM preparation. Brush-border SO4-OH exchange was further localized by demonstrating Na-stimulated SO4 efflux from vesicles loaded under pH gradient conditions, suggesting that Na-SO4 cotransport and SO4-OH exchange are on the same BBM vesicles. In conclusion, a SO4-OH exchanger (or H-SO4 cotransporter) exists on the brush border of rabbit ileum which is distinct from the brush-border Cl-HCO3(OH) exchanger.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

在存在pH梯度(内部为7.7,外部为5.5)的情况下,与不存在pH梯度时相比,兔回肠刷状缘膜(BBM)囊泡摄取SO4的初始速度显著加快。在pH梯度条件下,SO4会短暂积累,其浓度比平衡时高13倍(“超射现象”)。与单独的pH梯度相比,叠加HCO3梯度并不会进一步刺激SO4摄取的初始速度。表明这种pH梯度刺激的SO4摄取代表SO4-OH交换的证据包括:SO4转运对膜电位变化不敏感;阴离子交换抑制剂DIDS和SITS对SO4摄取有85%-95%的抑制作用;以及饱和动力学(SO4的Km = 0.475 +/- 0.054 mM;Vmax = 4.1 +/- 0.1 nmol SO4 X mg蛋白-1 X min-1)。硫酸盐不会抑制pH梯度刺激的36Cl摄取,这表明SO4-OH和Cl-HCO3(OH)是不同的交换体。当将BBM囊泡与基底外侧膜(BLM)囊泡进行比较时,发现pH梯度刺激的SO4摄取主要存在于BBM制剂中。通过证明在pH梯度条件下加载的囊泡中Na刺激的SO4流出,进一步确定了刷状缘SO4-OH交换的位置,这表明Na-SO4共转运和SO4-OH交换存在于同一BBM囊泡上。总之,兔回肠刷状缘存在一种SO4-OH交换体(或H-SO与刷状缘Cl-HCO3(OH)交换体不同。(摘要截短于250字)

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