TY - JOUR
T1 - Helical image reconstruction of the outward-open human erythrocyte band 3 membrane domain in tubular crystals
AU - Yamaguchi, Tomohiro
AU - Fujii, Takashi
AU - Abe, Yoshito
AU - Hirai, Teruhisa
AU - Kang, Dongchon
AU - Namba, Keiichi
AU - Hamasaki, Naotaka
AU - Mitsuoka, Kaoru
N1 - Funding Information:
We thank Yoshinori Fujiyoshi, Yoko Hiroaki, and Kazutoshi Tani for invaluable discussions. We also thank Kazumi Kobayashi (JEOL DATUM) for technical advice about cryo-electron microscopy. Xiu Ri Jin, Kayoko Tsunematsu, and Takako Ogata provided technical support during sample purification. This research was supported in part by Grants-in-Aid for Scientific Research on Priority Area ‘Membrane Interface’ to T.Y. and on Priority Area ‘Biological Macromolecular Assemblies’ to K.N. (16087207) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan; and by the Japan New Energy and Industrial Technology Development Organization (NEDO) to K.M.
PY - 2010/3
Y1 - 2010/3
N2 - The C-terminal membrane domain of erythrocyte band 3 functions as an anion exchanger. Here, we report the three-dimensional (3D) structure of the membrane domain in an inhibitor-stabilized, outward-open conformation at 18 Å resolution. Unstained, frozen-hydrated tubular crystals containing the membrane domain of band 3 purified from human red blood cells (hB3MD) were examined using cryo-electron microscopy and iterative helical real-space reconstruction (IHRSR). The 3D image reconstruction of the tubular crystals showed the molecular packing of hB3MD dimers with dimensions of 60 × 110 Å in the membrane plane and a thickness of 70 Å across the membrane. Immunoelectron microscopy and carboxyl-terminal digestion demonstrated that the intracellular surface of hB3MD was exposed on the outer surface of the tubular crystal. A 3D density map revealed that hB3MD consists of at least two subdomains and that the outward-open form is characterized by a large hollow area on the extracellular surface and continuous density on the intracellular surface.
AB - The C-terminal membrane domain of erythrocyte band 3 functions as an anion exchanger. Here, we report the three-dimensional (3D) structure of the membrane domain in an inhibitor-stabilized, outward-open conformation at 18 Å resolution. Unstained, frozen-hydrated tubular crystals containing the membrane domain of band 3 purified from human red blood cells (hB3MD) were examined using cryo-electron microscopy and iterative helical real-space reconstruction (IHRSR). The 3D image reconstruction of the tubular crystals showed the molecular packing of hB3MD dimers with dimensions of 60 × 110 Å in the membrane plane and a thickness of 70 Å across the membrane. Immunoelectron microscopy and carboxyl-terminal digestion demonstrated that the intracellular surface of hB3MD was exposed on the outer surface of the tubular crystal. A 3D density map revealed that hB3MD consists of at least two subdomains and that the outward-open form is characterized by a large hollow area on the extracellular surface and continuous density on the intracellular surface.
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U2 - 10.1016/j.jsb.2009.12.009
DO - 10.1016/j.jsb.2009.12.009
M3 - Article
C2 - 20005958
AN - SCOPUS:76749162402
VL - 169
SP - 406
EP - 412
JO - Journal of Structural Biology
JF - Journal of Structural Biology
SN - 1047-8477
IS - 3
ER -