Helical image reconstruction of the outward-open human erythrocyte band 3 membrane domain in tubular crystals

Tomohiro Yamaguchi, Takashi Fujii, Yoshito Abe, Teruhisa Hirai, Dongchon Kang, Keiichi Namba, Naotaka Hamasaki, Kaoru Mitsuoka

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)406-412
Number of pages7
JournalJournal of Structural Biology
Volume169
Issue number3
DOIs
Publication statusPublished - Mar 1 2010

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Computer-Assisted Image Processing
Erythrocytes
Membranes
Cryoelectron Microscopy
Immunoelectron Microscopy
Erythrocyte Membrane
Anions
Digestion

All Science Journal Classification (ASJC) codes

  • Structural Biology

Cite this

Helical image reconstruction of the outward-open human erythrocyte band 3 membrane domain in tubular crystals. / Yamaguchi, Tomohiro; Fujii, Takashi; Abe, Yoshito; Hirai, Teruhisa; Kang, Dongchon; Namba, Keiichi; Hamasaki, Naotaka; Mitsuoka, Kaoru.

In: Journal of Structural Biology, Vol. 169, No. 3, 01.03.2010, p. 406-412.

Research output: Contribution to journalArticle

Yamaguchi, Tomohiro ; Fujii, Takashi ; Abe, Yoshito ; Hirai, Teruhisa ; Kang, Dongchon ; Namba, Keiichi ; Hamasaki, Naotaka ; Mitsuoka, Kaoru. / Helical image reconstruction of the outward-open human erythrocyte band 3 membrane domain in tubular crystals. In: Journal of Structural Biology. 2010 ; Vol. 169, No. 3. pp. 406-412.
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AU - Hamasaki, Naotaka

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