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Title: A neutron crystallographic analysis of T{sub 6} porcine insulin at 2.1 Å resolution

Abstract

The charge balance and hydrogen-bonding network at the core of the insulin T{sub 6} hexamer have been investigated by neutron diffraction analysis at 2.1 Å resolution. Neutron diffraction data for T{sub 6} porcine insulin were collected to 2.1 Å resolution from a single crystal partly deuterated by exchange of mother liquor. A maximum-likelihood structure refinement was undertaken using the neutron data and the structure was refined to a residual of 0.179. The hydrogen-bonding network of the central core of the hexamer was observed and the charge balance between positively charged Zn ions and their surrounding structure was interpreted by considering the protonation and/or deprotonation states and interactions of HisB10, water and GluB13. The observed double conformation of GluB13 was essential to interpreting the charge balance and could be compared with the structure of a dried crystal of T{sub 6} human insulin at 100 K. Differences in the dynamic behaviour of the water molecules coordinating the upper and lower Zn ions were observed and interpreted. The hydrogen bonds in the insulin molecules, as well as those involving HisB10 and GluB13, are discussed. The hydrogen/deuterium (H/D) exchange ratios of the amide H atoms of T{sub 6} porcine insulin in crystals were obtainedmore » and showed that regions highly protected from H/D exchange are concentrated in the centre of a helical region of the B chains. From the viewpoint of soaking time versus H/D-exchange ratios, the amide H atoms can be classified into three categories.« less

Authors:
 [1];  [2];  [3];  [4];  [1];  [4];  [1]; ;  [3];  [2];  [5]
  1. Graduate School of Science and Engineering, Ibaraki University, Hitachi, Naka-Narusawa 4-12-1, Ibaraki 316-8511 (Japan)
  2. Frontier Research Center for Applied Atomic Sciences, Ibaraki University, Shirakata 162-1, Tokai, Ibaraki 319-1106 (Japan)
  3. Japan Atomic Energy Agency, Shirakata-shirane 2-4, Tokai, Ibaraki 319-1195 (Japan)
  4. Faculty of Engineering, Ibaraki University, Hitachi, Naka-Narusawa 4-12-1, Ibaraki 316-8511 (Japan)
  5. (Japan)
Publication Date:
OSTI Identifier:
22351189
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 65; Journal Issue: Pt 10; Other Information: PMCID: PMC2756163; PMID: 19770501; PUBLISHER-ID: be5122; OAI: oai:pubmedcentral.nih.gov:2756163; Copyright (c) International Union of Crystallography 2009; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; ATOMS; BONDING; CHAINS; DEUTERIUM; HYDROGEN; INTERACTIONS; MOLECULES; MONOCRYSTALS; NEUTRON DIFFRACTION; NEUTRONS; RESOLUTION; WATER

Citation Formats

Iwai, Wakari, Yamada, Taro, Kurihara, Kazuo, Ohnishi, Yuki, Kobayashi, Yoichiro, Tanaka, Ichiro, Takahashi, Haruyuki, Kuroki, Ryota, Tamada, Taro, Niimura, Nobuo, E-mail: niimura@mx.ibaraki.ac.jp, and Graduate School of Science and Engineering, Ibaraki University, Hitachi, Naka-Narusawa 4-12-1, Ibaraki 316-8511. A neutron crystallographic analysis of T{sub 6} porcine insulin at 2.1 Å resolution. Denmark: N. p., 2009. Web. doi:10.1107/S090744490902770X.
Iwai, Wakari, Yamada, Taro, Kurihara, Kazuo, Ohnishi, Yuki, Kobayashi, Yoichiro, Tanaka, Ichiro, Takahashi, Haruyuki, Kuroki, Ryota, Tamada, Taro, Niimura, Nobuo, E-mail: niimura@mx.ibaraki.ac.jp, & Graduate School of Science and Engineering, Ibaraki University, Hitachi, Naka-Narusawa 4-12-1, Ibaraki 316-8511. A neutron crystallographic analysis of T{sub 6} porcine insulin at 2.1 Å resolution. Denmark. doi:10.1107/S090744490902770X.
Iwai, Wakari, Yamada, Taro, Kurihara, Kazuo, Ohnishi, Yuki, Kobayashi, Yoichiro, Tanaka, Ichiro, Takahashi, Haruyuki, Kuroki, Ryota, Tamada, Taro, Niimura, Nobuo, E-mail: niimura@mx.ibaraki.ac.jp, and Graduate School of Science and Engineering, Ibaraki University, Hitachi, Naka-Narusawa 4-12-1, Ibaraki 316-8511. Thu . "A neutron crystallographic analysis of T{sub 6} porcine insulin at 2.1 Å resolution". Denmark. doi:10.1107/S090744490902770X.
@article{osti_22351189,
title = {A neutron crystallographic analysis of T{sub 6} porcine insulin at 2.1 Å resolution},
author = {Iwai, Wakari and Yamada, Taro and Kurihara, Kazuo and Ohnishi, Yuki and Kobayashi, Yoichiro and Tanaka, Ichiro and Takahashi, Haruyuki and Kuroki, Ryota and Tamada, Taro and Niimura, Nobuo, E-mail: niimura@mx.ibaraki.ac.jp and Graduate School of Science and Engineering, Ibaraki University, Hitachi, Naka-Narusawa 4-12-1, Ibaraki 316-8511},
abstractNote = {The charge balance and hydrogen-bonding network at the core of the insulin T{sub 6} hexamer have been investigated by neutron diffraction analysis at 2.1 Å resolution. Neutron diffraction data for T{sub 6} porcine insulin were collected to 2.1 Å resolution from a single crystal partly deuterated by exchange of mother liquor. A maximum-likelihood structure refinement was undertaken using the neutron data and the structure was refined to a residual of 0.179. The hydrogen-bonding network of the central core of the hexamer was observed and the charge balance between positively charged Zn ions and their surrounding structure was interpreted by considering the protonation and/or deprotonation states and interactions of HisB10, water and GluB13. The observed double conformation of GluB13 was essential to interpreting the charge balance and could be compared with the structure of a dried crystal of T{sub 6} human insulin at 100 K. Differences in the dynamic behaviour of the water molecules coordinating the upper and lower Zn ions were observed and interpreted. The hydrogen bonds in the insulin molecules, as well as those involving HisB10 and GluB13, are discussed. The hydrogen/deuterium (H/D) exchange ratios of the amide H atoms of T{sub 6} porcine insulin in crystals were obtained and showed that regions highly protected from H/D exchange are concentrated in the centre of a helical region of the B chains. From the viewpoint of soaking time versus H/D-exchange ratios, the amide H atoms can be classified into three categories.},
doi = {10.1107/S090744490902770X},
journal = {Acta Crystallographica. Section D: Biological Crystallography},
number = Pt 10,
volume = 65,
place = {Denmark},
year = {Thu Oct 01 00:00:00 EDT 2009},
month = {Thu Oct 01 00:00:00 EDT 2009}
}