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- Title
Charge-state-distribution analysis of Bach2 intrinsically disordered heme binding region.
- Authors
Tomoji Suenaga; Miki Watanabe-Matsui; Tamami Uejima; Hiroki Shima; Toshitaka Matsui; Masao Ikeda-Saito; Mikako Shirouzu; Kazuhiko Igarashi; Kazutaka Murayama
- Abstract
Bach2 is a transcriptional repressor that plays an important role in the differentiation of T-cells and B-cells. Bach2 is functionally regulated by heme binding, and possesses five Cys-Pro Cys-Pro (CP)-motifs as the heme binding site. To reveal the molecular mechanism of heme binding by Bach2, the intrinsically disordered heme binding region (a.a. 331 -520; Bach2331–520) and its CP-motif mutant were prepared and characterized with and without heme, by UV-Vis spectroscopy and thermal profiles. In addition, the charge-state-distributions (CSDs) were assessed by electrospray ionization mass spectrometry. The UV-Vis spectroscopy revealed a lack of five-coordinated heme binding in the CP-motif mutant of Bach2331–520. The thermal profile and CSDs of Bach2331–520 indicated that heme binding induces the destabilization of Bach2331–520. The thermal profile revealed that the wild type Bach2331–520 was destabilized more than the CP-motif mutant. The shift in the CSDs by heme binding suggested that heme binding causes Bach2331–520 to adopt a more compact conformation. In addition, heme binding to the CP-motif could reduce the flexibility of Bach2331–520. Consequently, the five-coordinated heme binding destabilizes Bach2331–520, by reducing the flexibility of the polypeptide chain.
- Subjects
HEME; BINDING sites; GENETIC repressors; CONFORMATIONAL analysis; ELECTROSPRAY ionization mass spectrometry
- Publication
Journal of Biochemistry, 2016, Vol 160, Issue 5, p291
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/jb/mvw035