基于受体亲和力的 PfEMP1 蛋白纯化。

Receptor Affinity-Based Purification of PfEMP1 Proteins.

机构信息

Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, Centre for Medical Parasitology, University of Copenhagen, Copenhagen, Denmark.

Department of Immunology, Noguchi Memorial Institute for Medical Research, University of Ghana, Legon, Ghana.

出版信息

Methods Mol Biol. 2022;2470:299-308. doi: 10.1007/978-1-0716-2189-9_22.

DOI:10.1007/978-1-0716-2189-9_22

PMID:35881354

Abstract

The virulence of Plasmodium falciparum is linked to the ability of infected erythrocytes (IEs) to bind a range of human receptors. This binding is mediated by a family of highly polymorphic proteins known as P. falciparum erythrocyte membrane protein 1 (PfEMP1). PfEMP1 proteins are expressed on the surface of IEs and are composed of extracellular domains (NTS, CIDR, DBL), a transmembrane region and an acidic C-terminal segment. Subdomains of the extracellular N-terminal part of PfEMP1 molecules have been shown to bind specific receptors.In this chapter, we describe how to purify PfEMP1 proteins by a receptor affinity-based method. This includes how to prepare affinity columns and how to subsequently test the functionality of the purified PfEMP1 protein in an ELISA-based assay.

摘要

疟原虫的毒力与感染的红细胞（IEs）结合一系列人类受体的能力有关。这种结合是由一系列高度多态性的蛋白质介导的，这些蛋白质被称为疟原虫红细胞膜蛋白 1（PfEMP1）。PfEMP1 蛋白在 IEs 的表面表达，由细胞外结构域（NTS、CIDR、DBL）、跨膜区和酸性 C 末端片段组成。PfEMP1 分子的细胞外 N 端部分的亚结构域已被证明与特定的受体结合。在本章中，我们描述了如何通过基于受体亲和力的方法来纯化 PfEMP1 蛋白。这包括如何制备亲和柱，以及如何随后在基于 ELISA 的测定中测试纯化的 PfEMP1 蛋白的功能。