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用于重组凝集素精氨酸取代的定向诱变破坏其细胞毒性活性。

Directed Mutagenesis for Arginine Substitution of a Recombinant Lectin Disrupts Its Cytotoxic Activity.

作者信息

Martínez-Alarcón Dania, Castro-Guillén José Luis, Fitches Elaine, Gatehouse John A, Przyborski Stefan, Moreno-Celis Ulisses, Blanco-Labra Alejandro, García-Gasca Teresa

机构信息

Centro de Investigación y de Estudios Avanzados Unidad Irapuato, Departamento de Biotecnología y Bioquímica, Irapuato 36821, Guanajuato, Mexico.

Tecnológico Nacional de México/Instituto Tecnológico Superior de Irapuato (ITESI), Km. 12.5, Carretera Irapuato-Silao, El Copal, Irapuato 36821, Guanajuato, Mexico.

出版信息

Int J Mol Sci. 2024 Dec 10;25(24):13258. doi: 10.3390/ijms252413258.

DOI:10.3390/ijms252413258
PMID:39769023
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11676905/
Abstract

Recently, we reported that a recombinant Tepary bean () lectin (rTBL-1) induces apoptosis in colon cancer cell lines and that cytotoxicity was related to differential recognition of β1-6 branched -glycans. Sequencing analysis and resolution of the rTBL-1 3D structure suggest that glycan specificity could be strongly influenced by two arginine residues, R103 and R130, located in the carbohydrate binding pocket. The aim of this work was to determine the contribution of these residues towards cytotoxic activity. Two rTBL-1 mutants were produced in , biochemically characterized, and cytotoxic effects were evaluated on human colorectal cancer cells (HT-29). Substitution of either of the arginine residues with glutamines resulted in significant reductions in cytotoxic activity, with losses of 1.5 and 3 times for R103 and R130, respectively. Docking analysis showed that the mutations decreased lectin affinity binding to some Epidermal Growth Factor Receptor (EGFR)-related -glycans. Together, these findings confirm that both of the selected arginine residues (R103 and R130) play a key role in the recognition of tumor cell glycoconjugates by rTBL-1.

摘要

最近，我们报道了一种重组 tepary 豆（）凝集素（rTBL-1）可诱导结肠癌细胞系凋亡，且细胞毒性与对β1-6 分支 -聚糖的差异识别有关。rTBL-1 的测序分析和三维结构解析表明，位于碳水化合物结合口袋中的两个精氨酸残基 R103 和 R130 可能会强烈影响聚糖特异性。这项工作的目的是确定这些残基对细胞毒性活性的贡献。在 中产生了两个 rTBL-1 突变体，对其进行了生化表征，并评估了它们对人结肠癌细胞（HT-29）的细胞毒性作用。用谷氨酰胺取代任何一个精氨酸残基都会导致细胞毒性活性显著降低，R103 和 R130 的活性损失分别为 1.5 倍和 3 倍。对接分析表明，这些突变降低了凝集素与一些表皮生长因子受体（EGFR）相关 -聚糖的亲和结合。总之，这些发现证实所选的两个精氨酸残基（R103 和 R130）在 rTBL-1 识别肿瘤细胞糖缀合物中起关键作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/c31192e229ea/ijms-25-13258-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/fbb77b791324/ijms-25-13258-g002.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/ad0fb2a244d0/ijms-25-13258-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/cfc1c6a5b00b/ijms-25-13258-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/ac3c843905e8/ijms-25-13258-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/93582c1b81ec/ijms-25-13258-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/c31192e229ea/ijms-25-13258-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/fbb77b791324/ijms-25-13258-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/76d1154ebdd3/ijms-25-13258-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/ad0fb2a244d0/ijms-25-13258-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/cfc1c6a5b00b/ijms-25-13258-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/ac3c843905e8/ijms-25-13258-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/93582c1b81ec/ijms-25-13258-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32a8/11676905/c31192e229ea/ijms-25-13258-g005.jpg

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