缺少一个H2A.H2B二聚体的核小体核心颗粒的制备及结构表征

Preparation and structural characterization of nucleosomal core particles lacking one H2A.H2B dimer.

作者信息

de la Escalera S, Nieto M A, Palacián E

机构信息

Centro de Biología Molecular, Universidad Autónoma de Madrid, Cantoblanco, Spain.

出版信息

Biochem Biophys Res Commun. 1988 Dec 15;157(2):541-7. doi: 10.1016/s0006-291x(88)80283-3.

DOI:10.1016/s0006-291x(88)80283-3

PMID:3202866

Abstract

Nucleosomal core particles lacking one H2A.H2B dimer, (H2A.H2B)1 (H3.H4)2/DNA (146 bp), have been prepared by treatment of nucleosomal cores with dimethylmaleic anhydride, a reversible reagent for protein amino groups. The preparative procedure is simple, produces quantitative conversion of nucleosomal cores into dimer-deficient cores without formation of other subnucleosomal particles, and can be applied to the preparation of different H2A.H2B-deficient mono and oligonucleosomal particles. The structural properties of the dimer-deficient cores and complete nucleosomal cores reconstituted from the deficient particles and H2A.H2B dimers have been studied by DNase I digestion, thermal denaturation and circular dichroism.

摘要

通过用二甲基马来酸酐（一种蛋白质氨基的可逆试剂）处理核小体核心，制备了缺少一个H2A.H2B二聚体的核小体核心颗粒，即(H2A.H2B)1 (H3.H4)2/DNA（146碱基对）。该制备过程简单，能将核小体核心定量转化为缺乏二聚体的核心，且不会形成其他亚核小体颗粒，还可用于制备不同的缺乏H2A.H2B的单核小体和寡核小体颗粒。通过DNase I消化、热变性和圆二色性研究了缺乏二聚体的核心以及由缺乏二聚体的颗粒和H2A.H2B二聚体重构的完整核小体核心的结构特性。