Kaltner Herbert, Solís Dolores, André Sabine, Lensch Martin, Manning Joachim C, Mürnseer Michael, Sáiz José Luis, Gabius Hans-Joachim
Institut fur Physiologische Chemie, Tierarztliche Fakultat, Ludwig-Maximilians-Universitat Munchen, Veterinarstrasse 13, D-80539 Munchen, Germany.
Biochemistry. 2009 May 26;48(20):4403-16. doi: 10.1021/bi900083q.
Animal galectins (lectins with specificity for beta-galactosides of glycan chains) are potent effectors in diverse aspects of cell sociology. Gene divergence has led to different groups and a marked interspecies variability in the number of members per group. Since the suitability of a model for studying functionality in the galectin network will be distinguished by a rather simple degree of complexity, we have focused on chicken galectins (CGs). Starting from partial expression sequence tag information, we here report on cloning of full-length cDNA for the first avian tandem-repeat-type galectin. It is termed CG-8 on the basis of its sequence similarity to galectin-8 from mammals. Systematic sequence searches revealed its unique character among CGs. Detection of two mature mRNA species points to production of isoforms. Alternative splicing affecting exon V generates the two proteins with linkers of either 9 (CG-8I) or 28 amino acids (CG-8II). Both proteins form monomers with a shape comparable to that of the proto-type proteins CG-1A/B in solution, act as cross-linkers in hemagglutination, and bind cells with a strict dependence on galactose. Western blotting revealed the presence of either CG-8II or the mixture in organ extracts. No evidence of a truncated form was obtained. Preparation of a specific antibody also enabled immunohistochemical localization. Prominent sites of its presence were defense cells in the l. propria mucosae, in addition to immune cells in distinct organs such as alveolar macrophages and thymocytes. Overall, we extend the network of CGs to a tandem-repeat-type protein and provide a detailed characterization from gene and protein structures to expression.
动物半乳糖凝集素(对聚糖链的β-半乳糖苷具有特异性的凝集素)是细胞社会学多个方面的有力效应物。基因分化导致了不同的组,且每组成员数量存在显著的种间差异。由于用于研究半乳糖凝集素网络功能的模型的适用性将由相当简单的复杂程度来区分,我们将重点放在了鸡半乳糖凝集素(CGs)上。从部分表达序列标签信息开始,我们在此报告首次克隆出禽串联重复型半乳糖凝集素的全长cDNA。基于其与哺乳动物半乳糖凝集素-8的序列相似性,将其命名为CG-8。系统的序列搜索揭示了它在CGs中的独特特征。检测到两种成熟的mRNA种类表明存在异构体。影响外显子V的可变剪接产生了两种蛋白质,其连接子分别为9个(CG-8I)或28个氨基酸(CG-8II)。两种蛋白质在溶液中均形成形状与原型蛋白质CG-1A/B相当的单体,在血细胞凝集中起交联剂作用,并且严格依赖半乳糖结合细胞。蛋白质印迹法显示器官提取物中存在CG-8II或其混合物。未获得截短形式的证据。制备特异性抗体也使得能够进行免疫组织化学定位。除了在不同器官如肺泡巨噬细胞和胸腺细胞中的免疫细胞外,其存在的主要部位是固有层黏膜中的防御细胞。总体而言,我们将CGs网络扩展到一种串联重复型蛋白质,并提供了从基因和蛋白质结构到表达的详细表征。