Ahmed Hafiz, Vasta Gerardo R
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 E. Pratt Street, Suite 236, Baltimore, MD 21202, USA.
Biochem Biophys Res Commun. 2008 Jul 4;371(3):350-5. doi: 10.1016/j.bbrc.2008.04.078. Epub 2008 Apr 28.
Galectins, a family of beta-galactoside-binding proteins, participate in a variety of biological processes, such as early development, tissue organization, immune regulation, and tumor evasion and metastasis. Although as many as fifteen bona fide galectins have been identified in mammals, but the detailed mechanisms of their biological roles still remain unclear for most. This fragmentary knowledge extends to galectin-like proteins such as the rat lens crystallin protein GRIFIN (Galectin-related inter fiber protein) and the galectin-related protein GRP (previously HSPC159; hematopoietic stem cell precursor) that lack carbohydrate-binding activity. Their inclusion in the galectin family has been debated, as they are considered products of evolutionary co-option. We have identified a homologue of the GRIFIN in zebrafish (Danio rerio) (designated DrGRIFIN), which like the mammalian equivalent is expressed in the lens, particularly in the fiber cells, as revealed by whole mount in situ hybridization and immunostaining of 2 dpf (days post fertilization) embryos. As evidenced by RT-PCR, it is weakly expressed in the embryos as early as 21 hpf (hour post fertilization) but strongly at all later stages tested (30 hpf and 3, 4, 6, and 7 dpf). In adult zebrafish tissues, however, DrGRIFIN is also expressed in oocytes, brain, and intestine. Unlike the mammalian homologue, DrGRIFIN contains all amino acids critical for binding to carbohydrate ligands and its activity was confirmed as the recombinant DrGRIFIN could be purified to homogeneity by affinity chromatography on a lactosyl-Sepharose column. Therefore, DrGRIFIN is a bona fide galectin family member that in addition to its carbohydrate-binding properties, may also function as a crystallin.
半乳糖凝集素是一类β-半乳糖苷结合蛋白,参与多种生物学过程,如早期发育、组织构建、免疫调节以及肿瘤逃逸和转移。尽管在哺乳动物中已鉴定出多达15种真正的半乳糖凝集素,但它们大多数生物学作用的详细机制仍不清楚。这种零碎的认识也延伸到了半乳糖凝集素样蛋白,如大鼠晶状体晶体蛋白GRIFIN(半乳糖凝集素相关纤维间蛋白)和缺乏碳水化合物结合活性的半乳糖凝集素相关蛋白GRP(以前称为HSPC159;造血干细胞前体)。它们是否属于半乳糖凝集素家族一直存在争议,因为它们被认为是进化共选择的产物。我们在斑马鱼(Danio rerio)中鉴定出了GRIFIN的一个同源物(命名为DrGRIFIN),通过对受精后2天(dpf)胚胎的整体原位杂交和免疫染色发现,与哺乳动物中的对应物一样,它在晶状体中表达,特别是在纤维细胞中。逆转录聚合酶链反应(RT-PCR)表明,它早在受精后21小时(hpf)就在胚胎中弱表达,但在所有后续测试阶段(30 hpf以及3、4、6和7 dpf)都强烈表达。然而,在成年斑马鱼组织中,DrGRIFIN也在卵母细胞、脑和肠道中表达。与哺乳动物同源物不同,DrGRIFIN包含所有与碳水化合物配体结合至关重要的氨基酸,并且其活性得到了证实,因为重组DrGRIFIN可以通过乳糖基琼脂糖柱上的亲和层析纯化至同质。因此,DrGRIFIN是一个真正的半乳糖凝集素家族成员,除了具有碳水化合物结合特性外,还可能作为晶体蛋白发挥作用。