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Chemical Inactivation of the E3 Ubiquitin Ligase Cereblon by Pomalidomide-based Homo-PROTACs
Published on: May 15, 2019
Successful acquisition of a neutralizing monoclonal antibody against a novel neutrophil-activating peptide,
Tatsuya Hattori1, Kenta Nakashima2, Takayuki Marutani2
1Laboratory of Peptide Science, Graduate School of Bio-Science, Nagahama Institute of Bio-Science and Technology, Nagahama, Shiga 526-0829, Japan; Laboratory of Protein Engineering, Graduate School of Bio-Science, Nagahama Institute of Bio-Science and Technology, Nagahama, Shiga 526-0829, Japan.
Abstract:
Mitocryptide-1 (MCT-1) is a novel neutrophil-activating peptide derived from mitochondrial cytochrome c oxidase subunit VIII, and its physiological role and involvement in various diseases have not yet been elucidated. Generating neutralizing antibodies against the function of MCT-1 is of particular importance for investigating its physiological and pathophysiological roles, because MCT-1 is a fragmented peptide of its mother protein and hence it is very difficult to manipulate its expression level genetically without affecting expression of the mother protein. Here, we report the successful generation of a neutralizing monoclonal antibody (MAb) against MCT-1. This MAb, designated NM1B1, which specifically bound to the region of positions 9-22 of MCT-1, showed concentration-dependent inhibition of MCT-1-induced migration and β-hexosaminidase release in neutrophilic/granulocytic differentiated HL-60 cells. Thus, NM1B1, as a neutralizing MAb against MCT-1, could elucidate not just the physiological regulatory mechanisms of MCT-1 but also its pathophysiological involvement in various inflammatory diseases in vivo.
Insights
Researchers developed a neutralizing antibody, NM1B1, targeting Mitocryptide-1 (MCT-1), a novel peptide involved in neutrophil activation. This antibody can help investigate MCT-1
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Mitocryptide-1 (MCT-1) is a novel neutrophil-activating peptide derived from mitochondrial cytochrome c oxidase subunit VIII.
- Its physiological role and involvement in diseases remain unclear.
- Genetic manipulation of MCT-1 is challenging due to its fragmented nature.
Purpose of the Study:
- To generate a neutralizing monoclonal antibody (MAb) against MCT-1.
- To investigate the physiological and pathophysiological roles of MCT-1.
Main Methods:
- Generation of a monoclonal antibody (MAb) against MCT-1.
- Characterization of the MAb's binding specificity and inhibitory function.
- Assay of MCT-1-induced neutrophil functions using differentiated HL-60 cells.
Main Results:
- Successful generation of a neutralizing MAb against MCT-1, designated NM1B1.
- NM1B1 specifically bound to positions 9-22 of MCT-1.
- NM1B1 demonstrated concentration-dependent inhibition of MCT-1-induced neutrophil migration and β-hexosaminidase release.
Conclusions:
- NM1B1 is a potent neutralizing antibody against MCT-1.
- This MAb can be a valuable tool for elucidating MCT-1's physiological regulatory mechanisms.
- NM1B1 may help investigate MCT-1's pathophysiological involvement in inflammatory diseases.
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