通过与裂褶多糖复合保护多核苷酸免受核酸酶介导的水解。

Protection of polynucleotides against nuclease-mediated hydrolysis by complexation with schizophyllan.

作者信息

Mizu Masami, Koumoto Kazuya, Kimura Taro, Sakurai Kazuo, Shinkai Seiji

机构信息

Department of Chemical Process and Environments, The University of Kitakyushu, 1-1, Hibikino, Wakamatu-ku, Kitakyushu, Fukuoka 808-0135, Japan.

出版信息

Biomaterials. 2004 Jul;25(15):3109-16. doi: 10.1016/j.biomaterials.2003.09.079.

DOI:10.1016/j.biomaterials.2003.09.079

PMID:14967545

Abstract

Schizophyllan is a beta-(1-->3)-D-glucan existing as a triple helix in water and as a single chain in dimethylsulfoxide (DMSO), respectively. As we already reported, when some homo-polynucleotide (for example, poly(dA) or poly(C)) is added to the schizophyllan/DMSO solution and subsequently DMSO is exchanged for water, the single chain of schizophyllan (s-SPG) forms a complex with the polynucleotide. The present work demonstrates that the polynucleotide bound in the complex is more stable to nuclease-mediated hydrolysis than the polynucleotide itself (i.e., naked polynucleotide), using high-performance liquid chromatography and ultraviolet absorbance technique. A kinetic study for the hydrolysis clarified that the simple Michaelis-Menten relation is held and the maximum velocity for the complex is one-sixth as small as that of the naked polynucleotide. This low hydrolysis rate for the complex suggests that s-SPG is applicable to a carrier for antisense oligonucleotides.

摘要

裂褶多糖是一种β-(1→3)-D-葡聚糖，在水中以三螺旋形式存在，在二甲基亚砜（DMSO）中以单链形式存在。正如我们之前报道的，当向裂褶多糖/DMSO溶液中加入一些同聚核苷酸（例如，聚(dA)或聚(C)），随后将DMSO换成水时，裂褶多糖单链（s-SPG）会与聚核苷酸形成复合物。本研究利用高效液相色谱和紫外吸收技术表明，复合物中结合的聚核苷酸比聚核苷酸本身（即裸聚核苷酸）对核酸酶介导的水解更稳定。对水解的动力学研究表明，符合简单的米氏关系，复合物的最大反应速度仅为裸聚核苷酸的六分之一。复合物这种较低的水解速率表明s-SPG可作为反义寡核苷酸的载体。