SEPARATION OF PHOSPHORYLATED HISTONE H1 VARIANTS BY HIGH-PERFORMANCE CAPILLARY ELECTROPHORESIS

被引：36

作者：

LINDNER, H ^{[1
]}

HELLIGER, W ^{[1
]}

DIRSCHLMAYER, A ^{[1
]}

TALASZ, H ^{[1
]}

WURM, M ^{[1
]}

SARG, B ^{[1
]}

JAQUEMAR, M ^{[1
]}

PUSCHENDORF, B ^{[1
]}

机构：

[1] BECKMAN INSTRUMENTS INC, A-3400 KLOSTERNEUBURG, AUSTRIA

来源：

JOURNAL OF CHROMATOGRAPHY | 1992年 / 608卷 / 1-2期

关键词：

D O I：

10.1016/0021-9673(92)87126-S

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

High-performance capillary electrophoresis (HPCE) was used to separate successfully distinct phosphorylated derivatives of individual histone H1 variants. With an untreated capillary (50 cm x 75 mum I.D.) the electrophoresis was performed in about 15 min. Inconvenient interactions of these highly basic proteins with the capillary wall were eliminated by using 0.1 M sodium phosphate buffer (pH 2.0) containing 0.03% hydroxypropylmethylcellulose. Under these experimental conditions the histone H1 variants H1b and H1c obtained from mitotic enriched NIH 3T3 fibroblasts and isolated by reversed-phase high-performance liquid chromatography were clearly separated in their non-phosphorylated and different phosphorylated forms. This result was confirmed by acid-urea gel electrophoresis, comparison with non-phosphorylated histones H1b and H1c, isolated from quiescent NIH 3T3 cells, and incubation of multi-phosphorylated histone Hlb with alkaline phosphatase and subsequent acid-urea and capillary electrophoresis. The results illustrate that the application of HPCE to the analysis of histone modifications provides a new alternative to traditional gel electrophoresis.

引用

页码：211 / 216

页数：6

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