In vitro expression and functional characterization of NPA motifs in aquaporins of Nosema bombycis
Autor: | Zhongyuan Shen, Ruisha Shang, Yiling Zhang, Jingru Qi, Shunming Tang, Zhilin Zhang, Gong Chen |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Mutant Xenopus Aquaporin Biology Aquaporins 03 medical and health sciences Xenopus laevis stomatognathic system Nosema Animals Amino Acid Sequence Conserved Sequence 030102 biochemistry & molecular biology General Veterinary Nosema bombycis Water General Medicine biology.organism_classification In vitro Cell biology 030104 developmental biology Infectious Diseases Water channel Insect Science Oocytes Parasitology Female Oligopeptides Biogenesis |
Zdroj: | Parasitology research. 117(11) |
ISSN: | 1432-1955 |
Popis: | Nosema bombycis contains functional aquaporins (NbAQPs), which are key targets for exploring the mechanism of N. bombycis infection; however, the regulation of these NbAQPs remains unknown. The two highly conserved asparagine-proline-alanine sequences (NPA motifs) play important roles in AQP biogenesis. As part of this study, we constructed a series of NbAQP mutants (NbAQP_NPA1, NbAQP_NPA2, and NbAQP_NPA1,2) and expressed them in BmN cells. The results showed that mutations in either NPA motif or in both NPA motifs did not affect NbAQP expression in vitro. After expression in Xenopus laevis oocytes, those injected with wild-type NbAQP rapidly expanded, whereas oocytes injected with NbAQP_NPAs did not significantly change in size. The associated water permeability (pf) of NbAQP_NPAs was significantly reduced five–six times compared to that of wild-type NbAQP. These results indicated that NPA motifs are necessary for the water channel function of AQPs in N. bombycis. The present study shows for the first time that the NbAQP NPA motif has an impact on the water permeability of aquaporin in N. bombycis, thereby providing a platform for further research into the mechanisms underlying the regulation of NbAQP expression. |
Databáze: | OpenAIRE |
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