Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel
Nicholas J Chandler
Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia; Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia
Ashleigh S Davey
Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia; Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia
Jonathan Y Weinstein
Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel
Julie V Nguyen
Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia
Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia; Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia
Ryan S Cross
Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia; Immunology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia
Misty R Jenkins
Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia; Immunology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia; La Trobe Institute of Molecular Science, La Trobe University, Bundoora, Victoria, Australia
Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia; Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia
Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia; Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia
De novo-designed receptor transmembrane domains (TMDs) present opportunities for precise control of cellular receptor functions. We developed a de novo design strategy for generating programmed membrane proteins (proMPs): single-pass α-helical TMDs that self-assemble through computationally defined and crystallographically validated interfaces. We used these proMPs to program specific oligomeric interactions into a chimeric antigen receptor (CAR) that we expressed in mouse primary T cells and found that both in vitro CAR T cell cytokine release and in vivo antitumor activity scaled linearly with the oligomeric state encoded by the receptor TMD, from monomers up to tetramers. All programmed CARs stimulated substantially lower T cell cytokine release relative to the commonly used CD28 TMD, which we show elevated cytokine release through lateral recruitment of the endogenous T cell costimulatory receptor CD28. Precise design using orthogonal and modular TMDs thus provides a new way to program receptor structure and predictably tune activity for basic or applied synthetic biology.
