Segmental isotope-labeling of the intrinsically disordered protein PQBP1

Yuko Nabeshima; Mineyuki Mizuguchi; Asagi Kajiyama; Hitoshi Okazawa

doi:10.1016/j.febslet.2014.10.028

Segmental isotope-labeling of the intrinsically disordered protein PQBP1

FEBS Lett. 2014 Dec 20;588(24):4583-9. doi: 10.1016/j.febslet.2014.10.028. Epub 2014 Nov 4.

Authors

Yuko Nabeshima¹, Mineyuki Mizuguchi², Asagi Kajiyama³, Hitoshi Okazawa⁴

Affiliations

¹ Faculty of Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan; Graduate School of Innovative Life Science, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan.
² Faculty of Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan; Graduate School of Innovative Life Science, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan. Electronic address: mineyuki@pha.u-toyama.ac.jp.
³ Faculty of Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan.
⁴ Department of Neuropathology, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; Center for Brain Integration Research, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.

PMID: 25447530
DOI: 10.1016/j.febslet.2014.10.028

Abstract

Polyglutamine tract-binding protein 1 (PQBP1) is an intrinsically disordered protein abundantly expressed in the brain. Mutations in the PQBP1 gene are causative for X-linked mental retardation disorders. Here, we investigated the structure of the C-terminal segment within the context of full-length PQBP1. We produced a segmentally isotope-labeled PQBP1 composed of a non-labeled segment (residues 1-219; N-segment) and a (13)C/(15)N-labeled segment (residues 220-265; C-segment). Our results demonstrate that the segmental isotope-labeling combined with NMR spectroscopy is useful for detecting a very weak intra-molecular interaction in an intrinsically disordered protein.

Keywords: Expressed protein ligation; Intra-molecular interaction; Intrinsically disordered protein; NMR; Segmental isotope-labeling.

Publication types

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

MeSH terms

Intrinsically Disordered Proteins / chemistry*
Intrinsically Disordered Proteins / metabolism*
Isotope Labeling*
Magnetic Resonance Spectroscopy
Nuclear Proteins / chemistry*
Nuclear Proteins / metabolism*

Substances

Intrinsically Disordered Proteins
Nuclear Proteins