Repurposing an endogenous degradation domain for antibody-mediated disposal of cell-surface proteins.

The exquisite specificity of antibodies can be harnessed to effect targeted degradation of membrane proteins. Here, we demonstrate targeted protein removal utilising a protein degradation domain derived from the endogenous human protein Proprotein Convertase Subtilisin/Kexin type 9 (PCSK9). Recombinant antibodies genetically fused to this domain drive the degradation of membrane proteins that undergo constitutive internalisation and recycling, including the transferrin receptor and the human cytomegalovirus latency-associated protein US28. We term this approach PACTAC (PCSK9-Antibody Clearance-Targeting Chimeras).

Description

Acknowledgements: This research was supported by the CIMR Flow Cytometry Core Facility. In particular, we wish to thank Gabriela Grondys-Kotarba and Reiner Schulte for their advice and support in flow cytometry and cell sorting. We also wish to thank Nika Romashova for her support in Cellomics microscopy. We would also like to thank Raimond Heukers, Timo De Groof, and Marine Smit for their advice on Vun100bv. BTK and DJO were supported by a Wellcome Trust Principal Research Fellowship to DJO (WT 207455/Z/17/Z). This work was supported by the Medical Research Council (grant numbers MR/S00081X/1) Programme Grant to J Sinclair and the Cambridge NIHR BRC Cell Phenotyping Hub. J Schmitt was supported by an MD Fellowship of the German Center for Infection Research (DZIF MD programme TI 07.003). SCG was supported by a Sir Henry Dale Fellowship, jointly funded by the Wellcome Trust and the Royal Society (098406/Z/12/B). For the purpose of open access, the authors have applied a Creative Commons Attribution (CC BY) license to any Author Accepted Manuscript version arising from this submission.

Funder: CIMR Flow Cytometry Core Facility

Funder: Cambridge NIHR BRC Cell Phenotyping Hub

Keywords

Human Cytomegalovirus, PACTAC, PCSK9, Targeted Protein Degradation, Humans, Proprotein Convertase 9, Serine Endopeptidases, Proprotein Convertases, Membrane Proteins, Receptors, LDL

Journal Title

EMBO Rep

Journal ISSN

1469-221X
1469-3178

Volume Title

25

Publisher

Springer Nature

Publisher DOI

https://doi.org/10.1038/s44319-024-00063-3

Rights and licensing

Except where otherwised noted, this item's license is described as Attribution 4.0 International

Sponsorship

Wellcome Trust (090909/Z/09/Z)
Medical Research Council (MR/S00081X/1)
Wellcome Trust (098406/Z/12/B)
Wellcome Trust (207455/Z/17/Z)
Wellcome Trust (098406/Z/12/Z)
Wellcome Trust (227915/Z/23/Z)

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