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选定 F 型凝集素的纯化和生化特性分析。

Purification and Biochemical Characterization of Selected F-Type Lectins.

机构信息

Department of Microbiology and Immunology, University of Maryland School of Medicine, UMB, and Institute of Marine and Environmental Technology, Columbus Center, Baltimore, MD, USA.

出版信息

Methods Mol Biol. 2020;2132:241-255. doi: 10.1007/978-1-0716-0430-4_24.

Abstract

The purification of fucose-binding lectins from the liver of striped bass (Morone saxatilis), a teleost fish, and the identification of a novel lectin sequence motif led to the identification of a new family of lectins, the F-type lectins (FTLs) (see overview of the FTL family in Chapter 23 ). Isolation and purification of these proteins from liver extracts of striped bass was accomplished by affinity chromatography and size exclusion, and their identification as FTLs, by direct Edman sequencing, and protein, transcript, and gene sequence analysis. The development of specific antibodies against the M. saxatilis FTL provided an additional tool for the identification of FTLs. These methods have been successfully used for the purification of the FTL family members from tissues and body fluids of various animal species. Production and characterization of FTLs has been facilitated by the expression of the recombinant proteins. In this chapter, the biochemical characterization of FTLs is focused on the analysis of their carbohydrate specificity.

摘要

从鲈形目鱼类条纹鲈(Morone saxatilis)的肝脏中纯化出岩藻糖结合凝集素,并鉴定出一种新型凝集素序列基序,这导致了一个新的凝集素家族的鉴定,即 F 型凝集素(FTLs)(见第 23 章对 FTL 家族的概述)。从条纹鲈肝脏提取物中通过亲和层析和大小排阻分离和纯化这些蛋白质,并通过直接 Edman 测序和蛋白质、转录物和基因序列分析将其鉴定为 FTL。针对 M. saxatilis FTL 的特异性抗体的开发为鉴定 FTL 提供了另一种工具。这些方法已成功用于从各种动物组织和体液中纯化 FTL 家族成员。通过表达重组蛋白,促进了 FTL 的生产和表征。在本章中,FTLs 的生化特性侧重于分析其碳水化合物特异性。

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