{"title":"受体酪氨酸激酶AXL中c -甘露糖基化的鉴定。","authors":"Kento Mori, Takehiro Suzuki, Urara Waki, Soichiro Hayashi, Shigehito Kadono, Ryota Kawahara, Minae Takeuchi, Hayato Mizuta, Naoshi Dohmae, Ryohei Katayama, Siro Simizu","doi":"10.1093/glycob/cwae096","DOIUrl":null,"url":null,"abstract":"<p><p>C-mannosylation is a unique type of glycosylation in which a mannose is added to tryptophan in a protein. However, the biological function of C-mannosylation is still largely unknown. AXL is a receptor tyrosine kinase, and its overexpression contributes to tumor malignancy. The role of AXL in cancer cells is broad, including invasion, drug resistance, and vasculogenic mimicry formation. Although Trp320 of AXL was predicted to be C-mannosylated, it has not been confirmed. Here, we demonstrated that Trp320 of AXL is C-mannosylated, measured by mass spectrometry of recombinant AXL purified from various cancer cells. Furthermore, re-expression of C-mannosylation-deficient AXL in human breast cancer MDA-MB-231 cells lacking AXL by the CRISPR/Cas9 system resulted in reduction of vasculogenic mimicry formation. Interestingly, phosphorylation levels of AKT in C-mannosylation-deficient AXL re-expressing cells were comparable to those of parental and wild-type AXL re-expressing cells. These results represent the first discovery of C-mannosylation in a receptor tyrosine kinase and the possibility that C-mannosylation may affect AXL function, distinct from its downstream signaling in cancer cells.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":"34 11","pages":""},"PeriodicalIF":3.4000,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11632359/pdf/","citationCount":"0","resultStr":"{\"title\":\"Identification of C-mannosylation in a receptor tyrosine kinase AXL.\",\"authors\":\"Kento Mori, Takehiro Suzuki, Urara Waki, Soichiro Hayashi, Shigehito Kadono, Ryota Kawahara, Minae Takeuchi, Hayato Mizuta, Naoshi Dohmae, Ryohei Katayama, Siro Simizu\",\"doi\":\"10.1093/glycob/cwae096\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>C-mannosylation is a unique type of glycosylation in which a mannose is added to tryptophan in a protein. However, the biological function of C-mannosylation is still largely unknown. AXL is a receptor tyrosine kinase, and its overexpression contributes to tumor malignancy. The role of AXL in cancer cells is broad, including invasion, drug resistance, and vasculogenic mimicry formation. Although Trp320 of AXL was predicted to be C-mannosylated, it has not been confirmed. Here, we demonstrated that Trp320 of AXL is C-mannosylated, measured by mass spectrometry of recombinant AXL purified from various cancer cells. Furthermore, re-expression of C-mannosylation-deficient AXL in human breast cancer MDA-MB-231 cells lacking AXL by the CRISPR/Cas9 system resulted in reduction of vasculogenic mimicry formation. Interestingly, phosphorylation levels of AKT in C-mannosylation-deficient AXL re-expressing cells were comparable to those of parental and wild-type AXL re-expressing cells. These results represent the first discovery of C-mannosylation in a receptor tyrosine kinase and the possibility that C-mannosylation may affect AXL function, distinct from its downstream signaling in cancer cells.</p>\",\"PeriodicalId\":12766,\"journal\":{\"name\":\"Glycobiology\",\"volume\":\"34 11\",\"pages\":\"\"},\"PeriodicalIF\":3.4000,\"publicationDate\":\"2024-09-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11632359/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Glycobiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1093/glycob/cwae096\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Glycobiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1093/glycob/cwae096","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
c -甘露糖基化是一种独特类型的糖基化,其中甘露糖被添加到蛋白质中的色氨酸中。然而,c -甘露糖基化的生物学功能在很大程度上仍然是未知的。AXL是一种酪氨酸激酶受体,其过表达与肿瘤恶性有关。AXL在癌细胞中的作用是广泛的,包括侵袭、耐药和血管生成模拟的形成。虽然AXL的Trp320被预测为c -甘露糖基化,但尚未得到证实。在这里,我们证明了AXL的Trp320是c -甘露糖基化的,通过从各种癌细胞中纯化的重组AXL的质谱测定。此外,通过CRISPR/Cas9系统在缺乏AXL的人乳腺癌MDA-MB-231细胞中重新表达缺乏c -甘露糖基化的AXL导致血管源性模拟形成减少。有趣的是,c -甘露糖基化缺陷AXL再表达细胞中AKT的磷酸化水平与亲代和野生型AXL再表达细胞相当。这些结果代表了受体酪氨酸激酶中c -甘露糖基化的首次发现,以及c -甘露糖基化可能影响AXL功能的可能性,不同于其在癌细胞中的下游信号传导。
Identification of C-mannosylation in a receptor tyrosine kinase AXL.
C-mannosylation is a unique type of glycosylation in which a mannose is added to tryptophan in a protein. However, the biological function of C-mannosylation is still largely unknown. AXL is a receptor tyrosine kinase, and its overexpression contributes to tumor malignancy. The role of AXL in cancer cells is broad, including invasion, drug resistance, and vasculogenic mimicry formation. Although Trp320 of AXL was predicted to be C-mannosylated, it has not been confirmed. Here, we demonstrated that Trp320 of AXL is C-mannosylated, measured by mass spectrometry of recombinant AXL purified from various cancer cells. Furthermore, re-expression of C-mannosylation-deficient AXL in human breast cancer MDA-MB-231 cells lacking AXL by the CRISPR/Cas9 system resulted in reduction of vasculogenic mimicry formation. Interestingly, phosphorylation levels of AKT in C-mannosylation-deficient AXL re-expressing cells were comparable to those of parental and wild-type AXL re-expressing cells. These results represent the first discovery of C-mannosylation in a receptor tyrosine kinase and the possibility that C-mannosylation may affect AXL function, distinct from its downstream signaling in cancer cells.
期刊介绍:
Established as the leading journal in the field, Glycobiology provides a unique forum dedicated to research into the biological functions of glycans, including glycoproteins, glycolipids, proteoglycans and free oligosaccharides, and on proteins that specifically interact with glycans (including lectins, glycosyltransferases, and glycosidases).
Glycobiology is essential reading for researchers in biomedicine, basic science, and the biotechnology industries. By providing a single forum, the journal aims to improve communication between glycobiologists working in different disciplines and to increase the overall visibility of the field.