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菠萝蜜种子中一种α-D-半乳糖特异性凝集素——桑科凝集素的结晶及初步X射线衍射研究。

Crystallization and preliminary X-ray diffraction studies of frutalin, an α-D-galactose-specific lectin from Artocarpus incisa seeds.

作者信息

Monteiro-Moreira Ana Cristina de Oliveira, D'Muniz Pereira Humberto, Vieira Neto Antonio Eufrasio, Mendes Batista Moreno Frederico Bruno, Duarte Pinto Lobo Marina, de Sousa Felipe Domingos, Moreira Renato de Azevedo

机构信息

Center of Experimental Biology (Nubex), University of Fortaleza (UNIFOR), 60811-905 Fortaleza-CE, Brazil.

Physics Institute of São Carlos, São Paulo University, Avenida Trabalhador São-carlense 400, Parque Arnold Schimidt, 13566-590 São Carlos-SP, Brazil.

出版信息

Acta Crystallogr F Struct Biol Commun. 2015 Oct;71(Pt 10):1282-5. doi: 10.1107/S2053230X15015186. Epub 2015 Sep 23.

Abstract

Frutalin is an α-D-galactose-specific carbohydrate-binding glycoprotein with antitumour properties and is a powerful tool for tumour biomarker discovery. The crystallization and preliminary X-ray diffraction analysis of this lectin, which was isolated from Artocarpus incisa seeds, are reported here. Frutalin was purified and submitted to mass-spectrometric analysis. Diverse masses at approximately 16 kDa were observed in the deconvoluted spectra, which support the presence of isoforms. The best frutalin crystals were grown within a week in 0.1 M citric acid pH 3.5 which contained 25% PEG 3350 as a precipitant at 293 K, and diffracted to a maximum resolution of 1.81 Å. The monoclinic crystals belonged to space group I2, with unit-cell parameters a = 76.17, b = 74.56, c = 118.98 Å, β = 96.56°. A molecular-replacement solution was obtained which indicated the presence of four monomers per asymmetric unit. Crystallographic refinement of the structure is in progress.

摘要

弗鲁他林是一种具有抗肿瘤特性的α-D-半乳糖特异性碳水化合物结合糖蛋白,是发现肿瘤生物标志物的有力工具。本文报道了从面包树种子中分离得到的这种凝集素的结晶及初步X射线衍射分析。弗鲁他林经过纯化并进行了质谱分析。在去卷积光谱中观察到约16 kDa处有多种质量数,这支持了同工型的存在。最佳的弗鲁他林晶体在293 K下,于含有25%聚乙二醇3350作为沉淀剂的0.1 M柠檬酸pH 3.5中一周内生长出来,其衍射极限分辨率为1.81 Å。单斜晶体属于空间群I2,晶胞参数a = 76.17,b = 74.56,c = 118.98 Å,β = 96.56°。获得了分子置换解,表明每个不对称单元存在四个单体。该结构的晶体学精修正在进行中。

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