دورية أكاديمية
Key residues for the light regulation of the blue light-activated adenylyl cyclase from Beggiatoa sp.
| العنوان: | Key residues for the light regulation of the blue light-activated adenylyl cyclase from Beggiatoa sp. |
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| المؤلفون: | Stierl M; Institut für Biologie/Experimentelle Biophysik, Humboldt Universität zu Berlin , Invalidenstrasse 42, D-10115 Berlin, Germany., Penzkofer A, Kennis JT, Hegemann P, Mathes T |
| المصدر: | Biochemistry [Biochemistry] 2014 Aug 12; Vol. 53 (31), pp. 5121-30. Date of Electronic Publication: 2014 Jul 28. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 0370623 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-4995 (Electronic) Linking ISSN: 00062960 NLM ISO Abbreviation: Biochemistry Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, American Chemical Society. |
| مواضيع طبية MeSH: | Adenylyl Cyclases/*chemistry , Adenylyl Cyclases/*radiation effects , Bacterial Proteins/*chemistry , Bacterial Proteins/*radiation effects , Beggiatoa/*enzymology , Photoreceptors, Microbial/*chemistry , Photoreceptors, Microbial/*radiation effects, Adenylyl Cyclases/genetics ; Amino Acid Substitution ; Bacterial Proteins/genetics ; Beggiatoa/genetics ; Beggiatoa/radiation effects ; Catalytic Domain ; Enzyme Activation/radiation effects ; Light ; Models, Molecular ; Mutagenesis, Site-Directed ; Optogenetics ; Photochemical Processes ; Photoreceptors, Microbial/genetics ; Protein Structure, Tertiary ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/radiation effects ; Signal Transduction |
| مستخلص: | Photoactivated adenylyl cyclases are powerful tools for optogenetics and for investigating signal transduction mechanisms in biological photoreceptors. Because of its large increase in enzyme activity in the light, the BLUF (blue light sensor using flavin adenine dinucleotide)-activated adenylyl cyclase (bPAC) from Beggiatoa sp. is a highly attractive model system for studying BLUF domain signaling. In this report, we studied the influence of site-directed mutations within the BLUF domain on the light regulation of the cyclase domain and determined key elements for signal transduction and color tuning. Photoactivation of the cyclase domain is accomplished via strand β5 of the BLUF domain and involves the formation of helical structures in the cyclase domain as assigned by vibrational spectroscopy. In agreement with earlier studies, we observed severely impaired signaling in mutations directly on strand β5 as well as in mutations affecting the hydrogen bond network around the flavin. Moreover, we identified a bPAC mutant with red-shifted absorbance and a decreased dark activity that is highly valuable for long-term optogenetic experiments. Additionally, we discovered a mutant that forms a stable neutral flavin semiquinone radical in the BLUF domain and surprisingly exhibits an inversion of light activation. |
| المشرفين على المادة: | 0 (Bacterial Proteins) 0 (Photoreceptors, Microbial) 0 (Recombinant Proteins) EC 4.6.1.1 (Adenylyl Cyclases) |
| تواريخ الأحداث: | Date Created: 20140722 Date Completed: 20141022 Latest Revision: 20151119 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1021/bi500479v |
| PMID: | 25046330 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 1520-4995 |
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| DOI: | 10.1021/bi500479v |