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膜核糖核酸酶和3'-核苷酸酶对大鼠肝细胞膜消化多聚尿苷酸的影响。

Effects of membrane ribonuclease and 3'-nucleotidase on the digestion of polyuridylic acid by rat liver plasma membrane.

作者信息

Aronson N N, Yannarell A

出版信息

Biochim Biophys Acta. 1975 Nov 17;413(1):135-42. doi: 10.1016/0005-2736(75)90064-4.

Abstract
  1. Fragments of isolated rat liver plasma membrane possess a ribonuclease activity which at pH 7.8 in the presence of 10 mM EDTA can digest polyuridylic acid (poly(U)) and polycytidylic acid (poly(C)) but not polyadenylic acid (poly(A)) and polyguanylic acid (poly(G)). Under these conditions, the membrane preparation does not degrade native or denatured DNA. 2. The products of the reaction with poly(U) (10 mM EDTA present) can be separated on DEAE-Sephadex into oligonucleotides of increasing chain length. Most of the products are di- to hexa-nucleotides which contain terminal 3'-phosphate groups. 3. When EDTA is not present (pH 7.8 or 8.8) the plasma membrane preparation degrades both poly(A) and poly(U). With poly(A) the product is all nucleoside while with poly(U) as substrate most of the product is nucleoside, but also some oligonucleotides are produced. 4. The ribonuclease releases acid soluble products very slowly from high concentrations of poly(U) (mg/ml). 5. Uridine trinucleotide with and without a terminal 3'-phosphate group is degraded by rat liver plasma membrane. The trinucleotide diphosphate is rapidly hydrolyzed to nucleoside while the trinucleotide itself is slowly digested and yields intermediate products, including nucleoside.
摘要
  1. 分离出的大鼠肝细胞膜碎片具有核糖核酸酶活性,在pH 7.8、存在10 mM乙二胺四乙酸(EDTA)的情况下,该酶能消化多聚尿苷酸(poly(U))和多聚胞苷酸(poly(C)),但不能消化多聚腺苷酸(poly(A))和多聚鸟苷酸(poly(G))。在这些条件下,膜制剂不会降解天然或变性的DNA。2. 与poly(U)反应的产物(存在10 mM EDTA)可在二乙氨基乙基葡聚糖(DEAE - Sephadex)上分离成链长不断增加的寡核苷酸。大多数产物是二至六核苷酸,其含有末端3'-磷酸基团。3. 当不存在EDTA时(pH 7.8或8.8),细胞膜制剂会降解poly(A)和poly(U)。以poly(A)为底物时,产物全是核苷;以poly(U)为底物时,大多数产物是核苷,但也会产生一些寡核苷酸。4. 核糖核酸酶从高浓度的poly(U)(mg/ml)中释放酸溶性产物的速度非常慢。5. 带有和不带有末端3'-磷酸基团的尿苷三磷酸均可被大鼠肝细胞膜降解。三磷酸二核苷酸迅速水解为核苷,而三磷酸本身则被缓慢消化并产生包括核苷在内的中间产物。

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