Pácha J, Teisinger J, Popp M, Capek K
Institute of Physiology, Czechoslovak Academy of Sciences, Vídenská.
J Membr Biol. 1991 Mar;120(3):201-10. doi: 10.1007/BF01868531.
Na,K-ATPase function was studied in order to evaluate the mechanism of increased colonic Na+ transport during early postnatal development. The maximum Na(+)-pumping activity that was represented by the equivalent short-circuit current after addition of nystatin (ISCN) did not change during postnatal life or after adrenalectomy performed in 16-day-old rats. ISCN was entirely inhibited by ouabain; the inhibitory constant was 0.1 mM in 10-day-old (young) and 0.4 mM in 90-day-old (adult) rats. The affinity of the Na,K pump for Na+ was higher in young (11 mM) than in adult animals (19 mM). The Na,K-ATPase activity (measured after unmasking of latent activity by treatment with sodium dodecylsulfate) increased during development and was also not influenced by adrenalectomy of 16-day-old rats. The inhibitory constant for ouabain (KI) was not changed during development (0.1-0.3 mM). Specific [3H]ouabain binding to isolated colonocytes increased during development (19 and 82 pmol/mg protein), the dissociation constant (KD) was 8 and 21 microM in young and adult rats, respectively. The Na+ turnover rate per single Na,K pump, which was calculated from ISCN and estimated density of binding sites per cm2 of tissue was 500 in adult and 6400 Na+/min.site in young rats. These data indicate that they very high Na+ transport during early postnatal life reflects an elevated turnover rate and increased affinity for Na+ of a single isoform of the Na,K pump. The development of Na+ extrusion across the basolateral membrane is not directly regulated by corticosteroids.
为了评估出生后早期发育过程中结肠钠转运增加的机制,对钠钾ATP酶的功能进行了研究。添加制霉菌素后以等效短路电流表示的最大钠泵活性(ISCN)在出生后或对16日龄大鼠进行肾上腺切除术后均未改变。ISCN完全被哇巴因抑制;在10日龄(幼年)大鼠中抑制常数为0.1 mM,在90日龄(成年)大鼠中为0.4 mM。幼年动物(11 mM)中钠钾泵对钠的亲和力高于成年动物(19 mM)。钠钾ATP酶活性(用十二烷基硫酸钠处理使潜在活性暴露后测量)在发育过程中增加,并且也不受16日龄大鼠肾上腺切除的影响。哇巴因的抑制常数(KI)在发育过程中未改变(0.1 - 0.3 mM)。发育过程中与分离的结肠细胞的特异性[3H]哇巴因结合增加(19和82 pmol/mg蛋白质),解离常数(KD)在幼年和成年大鼠中分别为8和21 μM。根据ISCN和每平方厘米组织结合位点的估计密度计算的每个钠钾泵的钠周转率在成年大鼠中为500,在幼年大鼠中为6400 Na+/分钟·位点。这些数据表明,出生后早期极高的钠转运反映了钠钾泵单一同工型的周转率升高和对钠的亲和力增加。基底外侧膜钠排泄的发育不受皮质类固醇的直接调节。