Open AccessDispatched uses Na+ flux to power release of lipid-modified Hedgehog
Author(s) -
Qianqian Wang,
Daniel Asarnow,
Ke Ding,
Randall K. Mann,
Jason Hatakeyama,
Yunxiao Zhang,
Yonghui Ma,
Yifan Cheng,
Philip A. Beachy
Publication year - 2021
Publication title -
nature
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 15.993
H-Index - 1226
eISSN - 1476-4687
pISSN - 0028-0836
DOI - 10.1038/s41586-021-03996-0
Subject(s) - transmembrane domain , transmembrane protein , hedgehog , chemistry , hedgehog signaling pathway , flux (metallurgy) , biophysics , patched , biology , biochemistry , amino acid , signal transduction , receptor , organic chemistry
The Dispatched protein, which is related to the NPC1 and PTCH1 cholesterol transporters 1,2 and to H + -driven transporters of the RND family 3,4 , enables tissue-patterning activity of the lipid-modified Hedgehog protein by releasing it from tightly -localized sites of embryonic expression 5-10 . Here we determine a cryo-electron microscopy structure of the mouse protein Dispatched homologue 1 (DISP1), revealing three Na + ions coordinated within a channel that traverses its transmembrane domain. We find that the rate of Hedgehog export is dependent on the Na + gradient across the plasma membrane. The transmembrane channel and Na + binding are disrupted in DISP1-NNN, a variant with asparagine substitutions for three intramembrane aspartate residues that each coordinate and neutralize the charge of one of the three Na + ions. DISP1-NNN and variants that disrupt single Na + sites retain binding to, but are impaired in export of the lipid-modified Hedgehog protein to the SCUBE2 acceptor. Interaction of the amino-terminal signalling domain of the Sonic hedgehog protein (ShhN) with DISP1 occurs via an extensive buried surface area and contacts with an extended furin-cleaved DISP1 arm. Variability analysis reveals that ShhN binding is restricted to one extreme of a continuous series of DISP1 conformations. The bound and unbound DISP1 conformations display distinct Na + -site occupancies, which suggests a mechanism by which transmembrane Na + flux may power extraction of the lipid-linked Hedgehog signal from the membrane. Na + -coordinating residues in DISP1 are conserved in PTCH1 and other metazoan RND family members, suggesting that Na + flux powers their conformationally driven activities.