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The immunotoxin activity of exotoxin A is sensitive to domain modifications

Author:
Amiri Tehranizadeh, Zeinab, Sankian, Mojtaba, Fazly Bazzaz, Bibi Sedigheh, Chamani, Jamshidkhan, Mehri, Soghra, Baratian, Ali, Saberi, Mohammad Reza
Source:
International journal of biological macromolecules 2019 v.134 pp. 1120-1131
ISSN:
0141-8130
Subject:
Pseudomonas aeruginosa, amino acids, antibodies, antineoplastic activity, clinical trials, exotoxins, humans, in vitro studies, molecular dynamics, neoplasms, point mutation, receptors, recombinant proteins, simulation models, solubility, solubilization, toxicity
Abstract:
Immunotoxins are a class of recombinant proteins which consist of an antibody and a part of a bacterial or herbal toxin. Immunotoxins containing Pseudomonas aeruginosa exotoxin A (PEA) have been found to be very applicable in clinical trials. Many obstacles such as solubility and absorbency reduce their usability in solid tumors. The current study aims to overcome the mentioned barriers by addition and removal of functional and non-functional domains with a structural approach. In the experimental section, we took advantage of molecular dynamics simulations to predict the functionality of candidate immunotoxins which target human HER2 receptors and confirmed our findings with in vitro experiments. We found out when no changes were made to domain II of PEA, addition of solubilizing domains to immunotoxins would not reduce their targeting and anti-tumor activity, while increasing the yield of expression and stability. On the other side, when we replaced domain II with eleven amino acids of furin cleavage site (FCS), the activity of the immunotoxin was mainly affected by the FCS neighboring domains and linkers. A combination of seven beneficial point mutations in domain III was also assessed and reconfirmed that the toxicity of the immunotoxin would be reduced dramatically.The obtained results indicate that the addition or removal of domains cannot depict the activity of immunotoxins and the matter should be assessed structurally in advance.
Agid:
6454693