Transferrin Mediated NCC Killing Activity through NCCRP-1 in Nile Tilapia (<i>Oreochromis niloticus</i>)
Non-specific cytotoxic cell l (NCC) is a kind of important lymphocyte participating in the non-specific immune response in teleost. Non-specific cytotoxic cell receptor protein 1 (NCCRP-1) is a receptor molecule on the surface of NCC and plays an important role in mediating the activity of NCC. Howe...
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MDPI AG
2022-09-01
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| Online Access: | https://www.mdpi.com/2410-3888/7/5/253 |
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| author | Yu Huang Zhengsi Chen Ruitao Xie Pei Wang Zhiqiang Zhang Jia Cai Bei Wang Jichang Jian |
| author_facet | Yu Huang Zhengsi Chen Ruitao Xie Pei Wang Zhiqiang Zhang Jia Cai Bei Wang Jichang Jian |
| author_sort | Yu Huang |
| collection | DOAJ |
| description | Non-specific cytotoxic cell l (NCC) is a kind of important lymphocyte participating in the non-specific immune response in teleost. Non-specific cytotoxic cell receptor protein 1 (NCCRP-1) is a receptor molecule on the surface of NCC and plays an important role in mediating the activity of NCC. However, there are few reports on which signal molecule could transmit signals through NCCRP-1. In this study, yeast two-hybrid library of tilapia liver and head kidney was constructed, and a transferrin from <i>Oreochromis niloticus</i> (On-TF) with interaction protein sequence was obtained by screening the library with bait vector NCCRP-1 of <i>Oreochromis niloticus</i> (On-NCCRP-1). Then, the open reading frame (ORF) of On-TF was cloned, which had 2088 bp in length, encoding polypeptides of 695 amino acids. The deduced amino acid sequence was highly homologous to teleost and similar to mammalian TF, containing two TR_FER domains (25-343th aa and 344-686th aa) for binding iron ions. Furthermore, a point-to-point yeast two-hybrid method was used to further verify whether On-NCCRP-1 could bind to On-TF. The recombinant On-TF (rOn-TF) protein was purified by prokaryotic expression system. In vitro experiments showed that rOn-TF could up-regulate the expression of killing effector molecule of NCC by On-NCCRP-1, and rOn-TF-activated NCCs showed a significantly improved ability to kill FHM cells, indicating that rOn-TF could regulate the NCC signaling pathway through NCC receptor molecule On-NCCRP-1. The results provide a more theoretical basis for understanding the regulation mechanism of NCC activity. |
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| language | English |
| last_indexed | 2024-03-09T20:13:34Z |
| publishDate | 2022-09-01 |
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| series | Fishes |
| spelling | doaj.art-6767d7df1a1049669601bc9606499e4b2023-11-24T00:06:17ZengMDPI AGFishes2410-38882022-09-017525310.3390/fishes7050253Transferrin Mediated NCC Killing Activity through NCCRP-1 in Nile Tilapia (<i>Oreochromis niloticus</i>)Yu Huang0Zhengsi Chen1Ruitao Xie2Pei Wang3Zhiqiang Zhang4Jia Cai5Bei Wang6Jichang Jian7Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, College of Fishery, Guangdong Ocean University, Zhanjiang 524000, ChinaGuangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, College of Fishery, Guangdong Ocean University, Zhanjiang 524000, ChinaKey Laboratory of Aquatic, Livestock, and Poultry Feed Science and Technology in South China, Ministry of Agriculture and Rural Affairs, Zhangjiang 524000, ChinaGuangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation, Beibu Gulf University, Qinzhou 535000, ChinaGuangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, College of Fishery, Guangdong Ocean University, Zhanjiang 524000, ChinaGuangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, College of Fishery, Guangdong Ocean University, Zhanjiang 524000, ChinaGuangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, College of Fishery, Guangdong Ocean University, Zhanjiang 524000, ChinaGuangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, College of Fishery, Guangdong Ocean University, Zhanjiang 524000, ChinaNon-specific cytotoxic cell l (NCC) is a kind of important lymphocyte participating in the non-specific immune response in teleost. Non-specific cytotoxic cell receptor protein 1 (NCCRP-1) is a receptor molecule on the surface of NCC and plays an important role in mediating the activity of NCC. However, there are few reports on which signal molecule could transmit signals through NCCRP-1. In this study, yeast two-hybrid library of tilapia liver and head kidney was constructed, and a transferrin from <i>Oreochromis niloticus</i> (On-TF) with interaction protein sequence was obtained by screening the library with bait vector NCCRP-1 of <i>Oreochromis niloticus</i> (On-NCCRP-1). Then, the open reading frame (ORF) of On-TF was cloned, which had 2088 bp in length, encoding polypeptides of 695 amino acids. The deduced amino acid sequence was highly homologous to teleost and similar to mammalian TF, containing two TR_FER domains (25-343th aa and 344-686th aa) for binding iron ions. Furthermore, a point-to-point yeast two-hybrid method was used to further verify whether On-NCCRP-1 could bind to On-TF. The recombinant On-TF (rOn-TF) protein was purified by prokaryotic expression system. In vitro experiments showed that rOn-TF could up-regulate the expression of killing effector molecule of NCC by On-NCCRP-1, and rOn-TF-activated NCCs showed a significantly improved ability to kill FHM cells, indicating that rOn-TF could regulate the NCC signaling pathway through NCC receptor molecule On-NCCRP-1. The results provide a more theoretical basis for understanding the regulation mechanism of NCC activity.https://www.mdpi.com/2410-3888/7/5/253aquaculturecytotoxicityimmune responsenatural killer cellapoptosis |
| spellingShingle | Yu Huang Zhengsi Chen Ruitao Xie Pei Wang Zhiqiang Zhang Jia Cai Bei Wang Jichang Jian Transferrin Mediated NCC Killing Activity through NCCRP-1 in Nile Tilapia (<i>Oreochromis niloticus</i>) Fishes aquaculture cytotoxicity immune response natural killer cell apoptosis |
| title | Transferrin Mediated NCC Killing Activity through NCCRP-1 in Nile Tilapia (<i>Oreochromis niloticus</i>) |
| title_full | Transferrin Mediated NCC Killing Activity through NCCRP-1 in Nile Tilapia (<i>Oreochromis niloticus</i>) |
| title_fullStr | Transferrin Mediated NCC Killing Activity through NCCRP-1 in Nile Tilapia (<i>Oreochromis niloticus</i>) |
| title_full_unstemmed | Transferrin Mediated NCC Killing Activity through NCCRP-1 in Nile Tilapia (<i>Oreochromis niloticus</i>) |
| title_short | Transferrin Mediated NCC Killing Activity through NCCRP-1 in Nile Tilapia (<i>Oreochromis niloticus</i>) |
| title_sort | transferrin mediated ncc killing activity through nccrp 1 in nile tilapia i oreochromis niloticus i |
| topic | aquaculture cytotoxicity immune response natural killer cell apoptosis |
| url | https://www.mdpi.com/2410-3888/7/5/253 |
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