Schubert Erik, Florin Nicole, Duthie Fraser, Henning Brewitz H, Kühl Toni, Imhof Diana, Hagelueken Gregor, Schiemann Olav
Institute of Physical and Theoretical Chemistry, University of Bonn, Wegelerstr. 12, D-53115 Bonn, Germany.
Pharmaceutical Chemistry I, Institute of Pharmacy, University of Bonn, Brühler Str. 7, D-53119 Bonn, Germany.
J Inorg Biochem. 2015 Jul;148:49-56. doi: 10.1016/j.jinorgbio.2015.05.008. Epub 2015 May 22.
The role of heme as a cofactor in enzymatic reactions has been studied for a long time and in great detail. Recently it was discovered that heme can also serve as a signalling molecule in cells but so far only few examples of this regulation have been studied. In order to discover new potentially heme-regulated proteins, we screened protein sequence databases for bacterial proteins that contain sequence features like a Cysteine-Proline (CP) motif, which is known for its heme-binding propensity. Based on this search we synthesized a series of these potential heme regulatory motifs (HRMs). We used cw EPR spectroscopy to investigate whether these sequences do indeed bind to heme and if the spin state of heme is changed upon interaction with the peptides. The corresponding proteins of two potential HRMs, FeoB and GlpF, were expressed and purified and their interaction with heme was studied by cw EPR and UV-Visible (UV-Vis) spectroscopy.
血红素作为酶促反应中的辅因子的作用已被长期且深入地研究。最近发现，血红素在细胞中还可作为信号分子，但到目前为止，仅对少数这种调节的例子进行了研究。为了发现新的潜在血红素调节蛋白，我们在蛋白质序列数据库中筛选了含有诸如半胱氨酸 - 脯氨酸（CP）基序等序列特征的细菌蛋白，该基序因其与血红素结合的倾向而闻名。基于此搜索，我们合成了一系列这些潜在的血红素调节基序（HRM）。我们使用连续波电子顺磁共振光谱（cw EPR光谱）来研究这些序列是否确实与血红素结合，以及血红素的自旋状态在与肽相互作用时是否发生变化。通过cw EPR和紫外 - 可见（UV - Vis）光谱对两种潜在HRM的相应蛋白FeoB和GlpF进行了表达和纯化，并研究了它们与血红素的相互作用。
