Expression and presentation of immune-related membrane proteins of fish by a cell surface display platform using insect cells
Autor: | Shuichi Asakawa, Yuzuru Suzuki, Satoshi Tasumi, Keisuke Kobayashi, Osamu Nakamura, Shihori Takanashi, Kiyoshi Kikuchi |
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Rok vydání: | 2019 |
Předmět: |
Fish Proteins
0301 basic medicine Insecta Immunology Cell Ligands Flow cytometry 03 medical and health sciences 0302 clinical medicine Immune system medicine Multiple cloning site Animals Receptors Immunologic Receptor Molecular Biology Cells Cultured medicine.diagnostic_test Chemistry Cell Membrane Fishes Lampreys Membrane Proteins Flow Cytometry Cell biology 030104 developmental biology medicine.anatomical_structure Membrane protein Cell culture Muramidase CC chemokine receptors Chickens 030215 immunology |
Zdroj: | Molecular Immunology. 114:553-560 |
ISSN: | 0161-5890 |
DOI: | 10.1016/j.molimm.2019.07.020 |
Popis: | Cell surface display is a useful platform to examine the interactions between two proteins of interest, such as immune receptors and ligands. This technique is also useful for studies on the immune receptors of lower vertebrates and invertebrates. However, in many cases, the commonly used cell culture temperature is relatively high for proteins from such organisms. Since insect cells can be cultured at lower temperatures than many other cells, and since they are equipped with “quality control” system, which is advantageous for the presentation of properly folded proteins, we anticipated that the insect cell surface display system could be more suitable for that type of research. In the present study, multiple cloning site of the commercially available expression vector pIB/V5-His was modified, and whether this vector could be useful to present fish immune-related membrane proteins was investigated. Using this plasmid, fugu’s CD8α and CC chemokine receptor 7 could be presented on the cell surface. The clones of the lamprey variable lymphocyte receptors obtained previously by the yeast surface display (YSD) system as hen’s egg lysozyme (HEL) binders also could be presented on the cell surface and bound to HEL. These results suggest that functional immune-related membrane proteins can be presented on the insect cell surface, indicating that this system is useful for immunological studies on exothermal animals. |
Databáze: | OpenAIRE |
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