Real time ligand-induced motion mappings of AChBP and nAChR using X-ray single molecule tracking

Hiroshi Sekiguchi; Yasuhito Suzuki; Yuri Nishino; Suzuko Kobayashi; Yoshiko Shimoyama; Weiyan Cai; Kenji Nagata; Masato Okada; Kouhei Ichiyanagi; Noboru Ohta; Naoto Yagi; Atsuo Miyazawa; Tai Kubo; Yuji C Sasaki

doi:10.1038/srep06384

Real time ligand-induced motion mappings of AChBP and nAChR using X-ray single molecule tracking

Sci Rep. 2014 Sep 16:4:6384. doi: 10.1038/srep06384.

Authors

Affiliations

¹ 1] CREST Sasaki Team, Japan Science and Technology Agency, The University of Tokyo, #609 Kiban Bldg., 5-1-5 Kashiwanoha, Kashiwa City, Chiba, 277-8561, Japan [2] Research &Utilization Division, Japan Synchrotron Radiation Research Institute, SPring-8 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5198, Japan.
² 1] CREST Sasaki Team, Japan Science and Technology Agency, The University of Tokyo, #609 Kiban Bldg., 5-1-5 Kashiwanoha, Kashiwa City, Chiba, 277-8561, Japan [2] Graduate School of Frontier Sciences, The University of Tokyo, Kiban Bldg., 5-1-5 Kashiwanoha, Kashiwa City, Chiba, 277-8561, Japan.
³ 1] Graduate School of Life Sciences, University of Hyogo, 3-2-1 Kouto, Kamigori-cho, Ako-gun, Hyogo, 679-1297, Japan [2] RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo, 679-5148, Japan.
⁴ Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
⁵ Graduate School of Frontier Sciences, The University of Tokyo, Kiban Bldg., 5-1-5 Kashiwanoha, Kashiwa City, Chiba, 277-8561, Japan.
⁶ Research &Utilization Division, Japan Synchrotron Radiation Research Institute, SPring-8 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5198, Japan.
⁷ 1] CREST Sasaki Team, Japan Science and Technology Agency, The University of Tokyo, #609 Kiban Bldg., 5-1-5 Kashiwanoha, Kashiwa City, Chiba, 277-8561, Japan [2] Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan [3] Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), 2-4-7 Aomi, Koto-ku, Tokyo 135-0064, Japan.
⁸ 1] CREST Sasaki Team, Japan Science and Technology Agency, The University of Tokyo, #609 Kiban Bldg., 5-1-5 Kashiwanoha, Kashiwa City, Chiba, 277-8561, Japan [2] Research &Utilization Division, Japan Synchrotron Radiation Research Institute, SPring-8 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5198, Japan [3] Graduate School of Frontier Sciences, The University of Tokyo, Kiban Bldg., 5-1-5 Kashiwanoha, Kashiwa City, Chiba, 277-8561, Japan.

Abstract

We observed the dynamic three-dimensional (3D) single molecule behaviour of acetylcholine-binding protein (AChBP) and nicotinic acetylcholine receptor (nAChR) using a single molecule tracking technique, diffracted X-ray tracking (DXT) with atomic scale and 100 μs time resolution. We found that the combined tilting and twisting motions of the proteins were enhanced upon acetylcholine (ACh) binding. We present the internal motion maps of AChBP and nAChR in the presence of either ACh or α-bungarotoxin (αBtx), with views from two rotational axes. Our findings indicate that specific motion patterns represented as biaxial angular motion maps are associated with channel function in real time and on an atomic scale.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acetylcholine / chemistry*
Acetylcholine / metabolism
Animals
Aplysia / growth & development
Aplysia / metabolism
Binding Sites
Bungarotoxins / chemistry*
Bungarotoxins / metabolism
Carrier Proteins / chemistry*
Carrier Proteins / metabolism
Crystallography, X-Ray
Models, Molecular
Protein Conformation
Receptors, Nicotinic / chemistry*
Receptors, Nicotinic / metabolism
Torpedo / growth & development
Torpedo / metabolism

Substances

Bungarotoxins
Carrier Proteins
Receptors, Nicotinic
Acetylcholine