دورية أكاديمية
Structure-Function Relationships in the Oligomeric NADPH-Dependent Assimilatory Sulfite Reductase.
| العنوان: | Structure-Function Relationships in the Oligomeric NADPH-Dependent Assimilatory Sulfite Reductase. |
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| المؤلفون: | Askenasy I, Murray DT, Andrews RM, Uversky VN; Department of Molecular Medicine and USF Health Byrd Alzheimer's Research Institute, Morsani College of Medicine , University of South Florida , Tampa , Florida 33612 , United States.; Institute for Biological Instrumentation of the Russian Academy of Sciences , Institutskaya strasse, 7 , Pushchino , Moscow Region 142290 , Russia., He H; Translational Science Laboratory, College of Medicine , Florida State University , Tallahassee , Florida 32306 , United States., Stroupe ME |
| المصدر: | Biochemistry [Biochemistry] 2018 Jul 03; Vol. 57 (26), pp. 3764-3772. Date of Electronic Publication: 2018 Jun 04. |
| نوع المنشور: | Journal Article; Research Support, U.S. Gov't, Non-P.H.S. |
| 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: | Escherichia coli/*metabolism , Sulfite Reductase (NADPH)/*metabolism, Amino Acid Sequence ; Escherichia coli/chemistry ; Intrinsically Disordered Proteins/chemistry ; Intrinsically Disordered Proteins/metabolism ; Models, Molecular ; NADP/metabolism ; Protein Binding ; Protein Conformation ; Protein Domains ; Protein Subunits/chemistry ; Protein Subunits/metabolism ; Sulfite Reductase (NADPH)/chemistry ; Thermodynamics |
| مستخلص: | The central step in the assimilation of sulfur is a six-electron reduction of sulfite to sulfide, catalyzed by the oxidoreductase NADPH-dependent assimilatory sulfite reductase (SiR). SiR is composed of two subunits. One is a multidomain flavin binding reductase (SiRFP) and the other an iron-containing oxidase (SiRHP). Both enzymes are primarily globular, as expected from their functions as redox enzymes. Consequently, we know a fair amount about their structures but not how they assemble. Curiously, both structures have conspicuous regions that are structurally undefined, leaving questions about their functions and raising the possibility that they are critical in forming the larger complex. Here, we used ultraviolet-visible and circular dichroism spectroscopy, isothermal titration calorimetry, proteolytic sensitivity tests, electrospray ionization mass spectrometry, and activity assays to explore the effect of altering specific amino acids in SiRFP on their function in the holoenzyme complex. Additionally, we used computational analysis to predict the propensity for intrinsic disorder within both subunits and found that SiRHP's N-terminus is predicted to have properties associated with intrinsic disorder. Both proteins also contained internal regions with properties indicative of intrinsic disorder. We showed that SiRHP's N-terminal disordered region is critical for complex formation. Together with our analysis of SiRFP amino acid variants, we show how molecular interactions outside the core of each SiR globular enzyme drive complex assembly of this prototypical oxidoreductase. |
| المشرفين على المادة: | 0 (Intrinsically Disordered Proteins) 0 (Protein Subunits) 53-59-8 (NADP) EC 1.8.1.2 (Sulfite Reductase (NADPH)) EC 1.8.1.2 (sulfite reductase (NADPH), E coli) |
| تواريخ الأحداث: | Date Created: 20180523 Date Completed: 20190220 Latest Revision: 20190320 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1021/acs.biochem.8b00446 |
| PMID: | 29787249 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 1520-4995 |
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| DOI: | 10.1021/acs.biochem.8b00446 |