副球菌素：其凝集素和酶学性质的纯化和验证。

Paracoccin: Purification and Validation of Its Lectin and Enzymatic Properties.

机构信息

Department of Cellular and Molecular Biology, Ribeirão Preto School of Medicine, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.

Laboratory of Immunochemistry and Glycobiology, Department of Cell and Molecular Biology and Pathogenic Bioagents, Ribeirão Preto School of Medicine University of São Paulo (FMRP/USP), Ribeirão Preto, SP, Brazil.

出版信息

Methods Mol Biol. 2020;2132:139-149. doi: 10.1007/978-1-0716-0430-4_14.

DOI:10.1007/978-1-0716-0430-4_14

PMID:32306322

Abstract

Studies on the effects of components derived from the human pathogenic fungi Paracoccidioides brasiliensis have identified paracoccin (PCN), as a bifunctional protein with lectin (GlcNAc-binding) and enzymatic (chitinase) activities, able to induce modulation of host immune response. Endogenous PCN acts as a fungal virulence factor, whereas exogenous purified PCN, administered to the host, confers protective immunity in a murine model of paracoccidioidomycosis. The immunomodulation induced by purified-PCN injection has characterized it as an agent applicable in the therapy and vaccine against paracoccidioidomycosis. This section describes methods for PCN purification and validation of its lectin and enzymatic activities. It includes detailed protocols to obtain homogeneous PCN from P. brasiliensis yeasts, as well as to purify recombinant PCN from transformed heterologous microorganisms.

摘要

从人类致病性真菌巴西副球孢子菌中提取的成分的研究已经鉴定出副球菌素 (PCN)，它是一种具有凝集素 (GlcNAc 结合) 和酶 (几丁质酶) 活性的双功能蛋白，能够诱导宿主免疫反应的调节。内源性 PCN 作为一种真菌毒力因子，而外源性纯化的 PCN，给予宿主，在巴西副球孢子菌病的小鼠模型中赋予保护免疫。纯化 PCN 注射诱导的免疫调节使其成为一种适用于巴西副球孢子菌病治疗和疫苗的药物。本节描述了 PCN 纯化方法及其凝集素和酶活性的验证。它包括从巴西副球孢子菌酵母中获得同源 PCN 以及从转化的异源微生物中纯化重组 PCN 的详细方案。