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TitleEffect of lipid head groups on double-layered two-dimensional crystals formed by aquaporin-0.
Journal, issue, pagesPLoS One, Vol. 10, Issue 1, Page e0117371, Year 2015
Publish dateJan 30, 2015
AuthorsRichard Kevin Hite / Po-Lin Chiu / Jan Michael Schuller / Thomas Walz /
PubMed AbstractAquaporin-0 (AQP0) is a lens-specific water channel that also forms membrane junctions. Reconstitution of AQP0 with dimyristoyl phosphatidylcholine (DMPC) and E. coli polar lipids (EPL) yielded well- ...Aquaporin-0 (AQP0) is a lens-specific water channel that also forms membrane junctions. Reconstitution of AQP0 with dimyristoyl phosphatidylcholine (DMPC) and E. coli polar lipids (EPL) yielded well-ordered, double-layered two-dimensional (2D) crystals that allowed electron crystallographic structure determination of the AQP0-mediated membrane junction. The interacting tetramers in the two crystalline layers are exactly in register, resulting in crystals with p422 symmetry. The high-resolution density maps also allowed modeling of the annular lipids surrounding the tetramers. Comparison of the DMPC and EPL bilayers suggested that the lipid head groups do not play an important role in the interaction of annular lipids with AQP0. We now reconstituted AQP0 with the anionic lipid dimyristoyl phosphatidylglycerol (DMPG), which yielded a mixture of 2D crystals with different symmetries. The different crystal symmetries result from shifts between the two crystalline layers, suggesting that the negatively charged PG head group destabilizes the interaction between the extracellular AQP0 surfaces. Reconstitution of AQP0 with dimyristoyl phosphatidylserine (DMPS), another anionic lipid, yielded crystals that had the usual p422 symmetry, but the crystals showed a pH-dependent tendency to stack through their cytoplasmic surfaces. Finally, AQP0 failed to reconstitute into membranes that were composed of more than 40% dimyristoyl phosphatidic acid (DMPA). Hence, although DMPG, DMPS, and DMPA are all negatively charged lipids, they have very different effects on AQP0 2D crystals, illustrating the importance of the specific lipid head group chemistry beyond its mere charge.
External linksPLoS One / PubMed:25635393 / PubMed Central
MethodsEM (electron crystallography)
Resolution7.0 - 8.0 Å
Structure data

EMDB-5802:
Electron crystallography of AQP0 with anionic phospholipids.
Method: EM (electron crystallography) / Resolution: 7.0 Å

EMDB-5803:
Electron crystallography of AQP0 with anionic phospholipids.
Method: EM (electron crystallography) / Resolution: 8.0 Å

Source
  • Ovis aries (sheep)

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