دورية أكاديمية
Protocol for detecting histidine polyphosphate modification of human proteins via MBP-tagged expression in E. coli.
| العنوان: | Protocol for detecting histidine polyphosphate modification of human proteins via MBP-tagged expression in E. coli. |
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| المؤلفون: | Lehotsky K; Department of Biomedical and Molecular Sciences, Queen's University, Kingston K7L 3N6, Canada., Neville N; Department of Biomedical and Molecular Sciences, Queen's University, Kingston K7L 3N6, Canada., Jia Z; Department of Biomedical and Molecular Sciences, Queen's University, Kingston K7L 3N6, Canada. Electronic address: jia@queensu.ca. |
| المصدر: | STAR protocols [STAR Protoc] 2024 Jun 21; Vol. 5 (2), pp. 102947. Date of Electronic Publication: 2024 Mar 11. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Cell Press Country of Publication: United States NLM ID: 101769501 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2666-1667 (Electronic) Linking ISSN: 26661667 NLM ISO Abbreviation: STAR Protoc Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: [Cambridge, MA] : Cell Press, [2020]- |
| مواضيع طبية MeSH: | Escherichia coli*/genetics , Escherichia coli*/metabolism , Polyphosphates*/metabolism , Polyphosphates*/chemistry , Maltose-Binding Proteins*/genetics , Maltose-Binding Proteins*/metabolism , Maltose-Binding Proteins*/chemistry , Histidine*/metabolism , Histidine*/genetics , Histidine*/chemistry , Protein Processing, Post-Translational*, Humans ; Blotting, Western |
| مستخلص: | Polyphosphate exhibits a unique post-translational modification-like function, known as histidine polyphosphate modification (HPM), marked by a robust non-covalent interaction with histidine repeat proteins. Here, we present a protocol for detecting HPM of human proteins via maltose-binding protein-tagged expression in E. coli. We describe steps for detecting HPM by observing electrophoretic mobility shifts on NuPAGE gels followed by western blot. We then detail procedures for analyzing the influence of ionic strength and pH on HPM. For complete details on the use and execution of this protocol, please refer to Neville et al. 1 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Molecular Biology; Protein Biochemistry; Protein expression and purification |
| المشرفين على المادة: | 0 (Polyphosphates) 0 (Maltose-Binding Proteins) 4QD397987E (Histidine) |
| تواريخ الأحداث: | Date Created: 20240312 Date Completed: 20240625 Latest Revision: 20240625 |
| رمز التحديث: | 20240625 |
| مُعرف محوري في PubMed: | PMC10943961 |
| DOI: | 10.1016/j.xpro.2024.102947 |
| PMID: | 38470910 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2666-1667 |
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| DOI: | 10.1016/j.xpro.2024.102947 |