tachylectin-2 的纯化和检测。

Purification and Assays of Tachylectin-2.

机构信息

Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan.

出版信息

Methods Mol Biol. 2020;2132:309-316. doi: 10.1007/978-1-0716-0430-4_30.

Abstract

Tachylectin-2, a 27-kDa protein consisting of a five-bladed β-propeller structure, is purified by three steps of chromatography, including dextran sulfate-Sepharose CL-6B, CM-Sepharose CL-6B, and Mono S. Three isolectins of tachylectin-2 including tachylectin-2a, -2b, and -2c are purified. These isolectins exhibit hemagglutinating activity against human A-type erythrocytes in a Ca-independent manner with tachylectin-2b showing the highest activity. Tachylectin-2b specifically agglutinates Staphylococcus saprophyticus KD. The tachylectin-2b-mediated hemagglutination is inhibited in the presence of GlcNAc and GalNAc. The association constants for GlcNAc and GalNAc are K = 1.95 × 10 M and K = 1.11 × 10 M, respectively. Ultracentrifugation analysis shows that tachylectin-2b is present in monomer form in solution.

摘要

速激肽素-2 是一种由五个叶片 β-桨叶结构组成的 27kDa 蛋白，通过葡聚糖硫酸盐-Sepharose CL-6B、CM-Sepharose CL-6B 和 Mono S 三步层析进行纯化。纯化得到三种速激肽素-2 的同工型，包括速激肽素-2a、-2b 和 -2c。这些同工型以 Ca 独立的方式对人 A 型红细胞表现出血凝活性，其中速激肽素-2b 表现出最高的活性。速激肽素-2b 特异性凝集腐生葡萄球菌 KD。在 GlcNAc 和 GalNAc 的存在下，速激肽素-2b 介导的血凝被抑制。GlcNAc 和 GalNAc 的缔合常数分别为 K = 1.95 × 10 M 和 K = 1.11 × 10 M。超速离心分析表明，速激肽素-2b 在溶液中以单体形式存在。