昆虫细胞表达的单纯疱疹病毒I型单链DNA结合蛋白ICP8的圆二色性分析

Circular dichroism analysis of insect cell expressed herpes simplex virus type I single-stranded DNA-binding protein ICP8.

作者信息

Spatz M, Ali S A, Auer M, Graf C, Eibl M M, Steinkasserer A

机构信息

Biomedizinische Forschungsgesellschaft mbH, c/o Institut für Bakteriologie, Mykologie und Hygiene, Veterinärmedizinische Universität Wien, Veterinaerplatz 1, Vienna, A-1210, Austria.

出版信息

Protein Expr Purif. 1999 Jun;16(1):40-6. doi: 10.1006/prep.1999.1046.

DOI:10.1006/prep.1999.1046

PMID:10336858

Abstract

We have developed a baculovirus expression system for the rapid and efficient production of large quantities (>5 mg/10(8) cells) of ICP8. The recombinant ICP8 is fully functional and binds to single-stranded DNA. Secondary structure calculations from circular dichroism measurements indicate a content of 34.5% alpha-helix and 15.4% beta-sheet. This is the first structural report for ICP8 using CD analysis, which will be very useful for high-throughput assay development and mechanistic studies.

摘要

我们已经开发出一种杆状病毒表达系统，用于快速高效地大量生产（>5毫克/10^8个细胞）ICP8。重组ICP8具有完全的功能，并且能与单链DNA结合。通过圆二色性测量进行的二级结构计算表明，其α-螺旋含量为34.5%，β-折叠含量为15.4%。这是首次使用圆二色性分析对ICP8进行结构报道，这对于高通量分析方法的开发和机理研究将非常有用。