دورية أكاديمية
Generation of 34 S-substituted protein-bound [4Fe-4S] clusters using 34 S-L-cysteine.
| العنوان: | Generation of 34 S-substituted protein-bound [4Fe-4S] clusters using 34 S-L-cysteine. |
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| المؤلفون: | Crack JC; Centre for Molecular and Structural Biochemistry, School of Chemistry, University of East Anglia, Norwich Research Park, Norwich, NR47 TJ, UK., Stewart MYY; Centre for Molecular and Structural Biochemistry, School of Chemistry, University of East Anglia, Norwich Research Park, Norwich, NR47 TJ, UK., Le Brun NE; Centre for Molecular and Structural Biochemistry, School of Chemistry, University of East Anglia, Norwich Research Park, Norwich, NR47 TJ, UK. |
| المصدر: | Biology methods & protocols [Biol Methods Protoc] 2019 Jan 22; Vol. 4 (1), pp. bpy015. Date of Electronic Publication: 2019 Jan 22 (Print Publication: 2019). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Oxford University Press Country of Publication: England NLM ID: 101693064 Publication Model: eCollection Cited Medium: Internet ISSN: 2396-8923 (Electronic) Linking ISSN: 23968923 NLM ISO Abbreviation: Biol Methods Protoc Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: [Oxford] : Oxford University Press, [2016]- |
| مستخلص: | The ability to specifically label the sulphide ions of protein-bound iron-sulphur (FeS) clusters with 34 S isotope greatly facilitates structure-function studies. In particular, it provides insight when using either spectroscopic techniques that probe cluster-associated vibrations, or non-denaturing mass spectrometry, where the ∼+2 Da average increase per sulphide enables unambiguous assignment of the FeS cluster and, where relevant, its conversion/degradation products. Here, we employ a thermostable homologue of the O -acetyl-l-serine sulfhydrylase CysK to generate 34 S-substituted l-cysteine and subsequently use it as a substrate for the l-cysteine desulfurase NifS to gradually supply 34 S 2- for in vitro FeS cluster assembly in an otherwise standard cluster reconstitution protocol. (© The Author(s) 2019. Published by Oxford University Press.) |
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| معلومات مُعتمدة: | BB/G019347/1 United Kingdom BB_ Biotechnology and Biological Sciences Research Council |
| فهرسة مساهمة: | Keywords: cysteine; iron–sulphur; mass spectrometry; resonance Raman spectroscopy |
| تواريخ الأحداث: | Date Created: 20200513 Latest Revision: 20210110 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC7200944 |
| DOI: | 10.1093/biomethods/bpy015 |
| PMID: | 32395620 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2396-8923 |
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| DOI: | 10.1093/biomethods/bpy015 |