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In the process of producing the recombinant pig Complement C5a anaphylatoxin (C5), yeast cells are transfected with a DNA expression vector containing the gene for the C5 protein (1-74aa) along with the N-terminal 6xHis-tag gene. Subsequent culturing induces the expression of the intended protein. The recombinant pig C5 protein is then collected and purified from the cell lysate through affinity purification, displaying a purity exceeding 90%, as verified by SDS-PAGE.
Complement C5a anaphylatoxin is a crucial protein involved in the inflammatory response and immune system regulation. It is generated by the cleavage of complement component C5. C5a plays diverse roles in various biological functions, extending beyond its classical role as an anaphylatoxin. It has been shown to have adverse effects on survival in sepsis and is implicated in critical illness-induced organ dysfunction [1][2][3][4]. Additionally, C5a has been demonstrated to exacerbate anaphylaxis and amplify mast cell activities, further underlining its significant role in immune responses [5][6]. C5a exerts neuroprotective effects and may have multiple effects relevant to neuronal survival [7].
Research has shown that C5a functions as a master switch for the pH balance in neutrophils, exerting fundamental immune metabolic effects and contributing to the overwhelming inflammatory response in sepsis [8]. Evidence has revealed that C5 blockade synergistically protects against lung cancer growth and metastasis [9]. Moreover, C5a exerts its role by binding to its receptor, CD88, after central nervous system injury, highlighting its involvement in the response to spinal cord injuries [10].
References:
[1] D. Rittirsch, M. Flierl, B. Nadeau, D. Day, M. Huber‐Lang, C. Mackayet al., Functional roles for c5a receptors in sepsis, Nature Medicine, vol. 14, no. 5, p. 551-557, 2008. https://doi.org/10.1038/nm1753
[2] A. Wood, A. Vassallo, C. Summers, E. Chilvers, & A. Morris, C5a anaphylatoxin and its role in critical illness‐induced organ dysfunction, European Journal of Clinical Investigation, vol. 48, no. 12, 2018. https://doi.org/10.1111/eci.13028
[3] R. O’Brien, N. Lynam‐Lennon, & M. Olcina, Thinking inside the box: intracellular roles for complement system proteins come into focus, British Journal of Cancer, vol. 128, no. 2, p. 165-167, 2023. https://doi.org/10.1038/s41416-022-02116-7
[4] P. Ward, Role of the complement in experimental sepsis, Journal of Leukocyte Biology, vol. 83, no. 3, p. 467-470, 2007. https://doi.org/10.1189/jlb.0607376
[5] M. Yasuda, Y. Tanaka, K. Bando, S. Sugawara, & K. Mizuta, Lipopolysaccharide priming exacerbates anaphylatoxin c5a-induced anaphylaxis in mice, Biological and Pharmaceutical Bulletin, vol. 46, no. 3, p. 432-439, 2023. https://doi.org/10.1248/bpb.b22-00766
[6] P. West, R. Bahri, K. Garcia-Rodriguez, G. Sweetland, G. Wileman, Á. Monteroet al., Interleukin-33 amplifies human mast cell activities induced by complement anaphylatoxins, Frontiers in Immunology, vol. 11, 2021. https://doi.org/10.3389/fimmu.2020.615236
[7] P. Mukherjee and G. Pasinetti, Complement anaphylatoxin c5a neuroprotects through mitogen‐activated protein kinase‐dependent inhibition of caspase 3, Journal of Neurochemistry, vol. 77, no. 1, p. 43-49, 2001. https://doi.org/10.1046/j.1471-4159.2001.00167.x
[8] S. Denk, M. Neher, D. Messerer, R. Wiegner, B. Nilsson, D. Rittirschet al., Complement c5a functions as a master switch for the ph balance in neutrophils exerting fundamental immunometabolic effects, The Journal of Immunology, vol. 198, no. 12, p. 4846-4854, 2017. https://doi.org/10.4049/jimmunol.1700393
[9] D. Ajona, S. Ortiz‐Espinosa, H. Moreno, T. Lozano, M. Pajares, J. Agorretaet al., A combined pd-1/c5a blockade synergistically protects against lung cancer growth and metastasis, Cancer Discovery, vol. 7, no. 7, p. 694-703, 2017. https://doi.org/10.1158/2159-8290.cd-16-1184
[10] Q. Guo, J. Cheng, H. Zhang, J. Zhang, B. Su, C. Bianet al., Expressions of c5a and its receptor cd88 after spinal cord injury in c3‐deficient mice, Scandinavian Journal of Immunology, vol. 77, no. 4, p. 224-229, 2013. https://doi.org/10.1111/sji.12001
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STRING: 9823.ENSSSCP00000005908
UniGene: Ssc.21108