Pyroglutamation of cell surface proteins CD47 and TRP1 by glutaminyl cyclase modulates therapeutic antibody binding
Anneloes van der Plas-van Duijn1,2, Levi M T Winkelman3, Claire Lensen1
1Department of Dermatology, Leiden University Medical Center, Leiden, the Netherlands.
Abstract:
Monoclonal antibodies are important modalities in the treatment of cancer. Post-translational modifications of proteins, such as glycosylation, can affect the binding affinity of therapeutic antibodies. Whether other PTMs modulate therapeutic antibody binding to different surface proteins is currently underexplored. Pyroglutamation is the post-translational cyclization of an N-terminal glutamine or glutamic acid residue into a pyroglutamate by glutaminyl cyclase. In this study, we investigated the impact of pyroglutamation on the binding affinity of three therapeutic antibodies targeting CD47 and TRP1. Here, we show that pyroglutamation on CD47 and TRP1 modulates the binding of anti(α)-CD47 magrolimab and αTRP1 TA99 and flanvotumab. Furthermore, the N-terminal glutamine on CD47 is crucial for effective antibody recognition, while pyroglutamation of TRP1 is involved in trafficking to the cell surface. These findings highlight that the pyroglutamation by glutaminyl cyclase can modulate the binding affinity of antibodies with therapeutic potential.
Insights
Pyroglutamation, a protein modification, impacts therapeutic antibody binding to cancer targets CD47 and TRP1. This modification affects antibody affinity and protein trafficking, highlighting its role in antibody-based cancer therapies.
Area of Science:
- Biochemistry
- Immunology
- Oncology
Background:
- Therapeutic monoclonal antibodies are crucial in cancer treatment.
- Post-translational modifications (PTMs) like glycosylation influence antibody binding.
- The effect of other PTMs, such as pyroglutamation, on therapeutic antibody binding is understudied.
Purpose of the Study:
- To investigate the impact of pyroglutamation on the binding affinity of therapeutic antibodies targeting CD47 and TRP1.
- To understand how pyroglutamation affects antibody recognition and protein localization.
Main Methods:
- Utilized pyroglutamation as a post-translational modification.
- Investigated binding affinity of therapeutic antibodies targeting CD47 and TRP1.
- Analyzed the role of N-terminal glutamine and pyroglutamation in antibody recognition and cell surface trafficking.
Main Results:
- Pyroglutamation of CD47 and TRP1 was shown to modulate the binding of anti-CD47 magrolimab and anti-TRP1 antibodies (TA99 and flanvotumab).
- N-terminal glutamine on CD47 is essential for antibody recognition.
- Pyroglutamation of TRP1 influences its trafficking to the cell surface.
Conclusions:
- Pyroglutamation by glutaminyl cyclase can alter the binding affinity of therapeutic antibodies.
- Understanding pyroglutamation's role is vital for developing effective antibody-based cancer therapies.
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