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RESEARCH | REPORT

S16B). As in the case of the C2 design, the C4 and diverse oligomeric states—are substantial 23. X. Feng, P. Barth, Nat. Chem. Biol. 12, 167–173 (2016).
symmetry axis of the design coincides with steps toward the complexity of natural trans- 24. A. D. Meruelo, S. K. Han, S. Kim, J. U. Bowie, Protein Sci. 21,
1746–1753 (2012).
the crystallographic axes of the crystal lattice membrane proteins with multiple membrane- 25. Y. Shi, Cell 159, 995–1014 (2014).
(fig. S16C). The four tetramer structures on the spanning regions and extra-membrane domains 26. R. Fernandez-Leiro, S. H. W. Scheres, Nature 537, 339–346
crystal C4 axes have overall structures very that play important roles in ligand/substrate (2016).
similar to each other and to the design model recognition and structure stabilization, such
(Fig. 4, F and G, and fig. S16A); the tetrameric as in the adenosine 5′-triphosphate–binding cas- ACKNOWLEDGMENTS
transmembrane domain, HR domain, and over- sette transporters, ion channels, ryanodine re- We thank J. Sumida for AUC support; A. Kang for crystallization
support; D. Ma and Z. Wang for crystallography support; and
all tetramer structure have Ca RMSDs to the de- ceptor, and g-secretase (25, 26). The capability to the staff at the Advanced Light Source and P. Huang, Y. Hsia,
sign model of 1.3 to 1.5 Å, 3.3 to 3.8 Å, and 3.3 to accurately design complex multipass transmem- A. Ford, L. Stewart, C. Xu, and many other members of the Baker
3.8 Å, respectively (Fig. 4H and fig. S16D, left). brane proteins that can be expressed in cells opens laboratory for helpful discussions. This work was facilitated by
The deviation in the HR domain may result the door to the design of a new generation of multi- the Hyak supercomputer at the University of Washington. Funding:
This work was supported by the Howard Hughes Medical
from crystal packing interactions between the pass membrane protein structures and functions. Institute (D.B.) and the National Institutes of Health (grant
termini; the Ca RMSDs over the first 162 resi- R01GM063919 to J.U.B.). P.L. was supported by the Raymond
dues are 2.2 to 2.3 Å (fig. S16D, right). The main and Beverly Sackler fellowship. D.M. was supported by the Basic
deviation from the design model is a tilting of REFERENCES AND NOTES Science Research Program through the National Research
the outer helices of transmembrane hairpins from 1. P.-S. Huang, S. E. Boyken, D. Baker, Nature 537, 320–327 Foundation of Korea funded by the Ministry of Education (grant
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Table S1
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