دورية أكاديمية
Heterogenous Expression and Purification of Lipid II Flippase from Staphylococcus aureus .
| العنوان: | Heterogenous Expression and Purification of Lipid II Flippase from Staphylococcus aureus . |
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| المؤلفون: | Zheng YY; State Key Laboratory of Chemical Biology and Drug Discovery, Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom, Hong Kong, China., Chung WH; State Key Laboratory of Chemical Biology and Drug Discovery, Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom, Hong Kong, China., Leung YC; State Key Laboratory of Chemical Biology and Drug Discovery, Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom, Hong Kong, China., Wong KY; State Key Laboratory of Chemical Biology and Drug Discovery, Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom, Hong Kong, China. |
| المصدر: | Protein and peptide letters [Protein Pept Lett] 2024; Vol. 31 (5), pp. e040724231578. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Bentham Science Publishers Country of Publication: Netherlands NLM ID: 9441434 Publication Model: Print Cited Medium: Internet ISSN: 1875-5305 (Electronic) Linking ISSN: 09298665 NLM ISO Abbreviation: Protein Pept Lett Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Hilversum, The Netherlands : Bentham Science Publishers Original Publication: Schiphol, The Netherlands : Bentham Science Publishers, c1994- |
| مواضيع طبية MeSH: | Staphylococcus aureus*/enzymology , Staphylococcus aureus*/genetics , Bacterial Proteins*/genetics , Bacterial Proteins*/chemistry , Bacterial Proteins*/isolation & purification , Bacterial Proteins*/biosynthesis , Bacterial Proteins*/metabolism, Detergents/chemistry ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Solubility ; Gene Expression ; Uridine Diphosphate N-Acetylmuramic Acid/analogs & derivatives |
| مستخلص: | Background: Staphylococcus aureus is a common pathogen with strains that are resistant to existing antibiotics. MurJ from S. aureus (SaMurJ), an integral membrane protein functioning as Lipid II flippase, is a potential target for developing new antibacterial agents against this pathogen. Successful expression and purification of this protein shall be useful in the development of drugs against this target. Objective: In this study, we demonstrated the optimized expression and purification procedures of SaMurJ, identified suitable detergent for extracting and solubilizing the protein, and examined the peptidisc system to generate a detergent-free environment. Methods: SaMurJ fused with N-terminal ten-His tag was expressed without induction. Six detergents were selected for screening the most efficient candidate for extraction and solubilization of the protein. The thermostability of the detergent-solubilized protein was assessed by evaluated temperature incubation. Different ratios of peptidisc bi-helical peptide (NSPr) to SaMurJ were mixed and the on-bead peptidisc assembly method was applied. Results: SaMurJ expressed in BL21(DE3) was confirmed by peptide fingerprinting, with a yield of 1 mg SaMurJ per liter culture. DDM was identified as the optimum detergent for solubilization and the nickel affinity column enabled SaMurJ purification with a purity of ~88%. However, NSPr could not stabilize SaMurJ. Conclusion: The expression and purification of SaMurJ were successful, with high purity and good yield. SaMurJ can be solubilized and stabilized by a DDM-containing buffer. (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.) |
| فهرسة مساهمة: | Keywords: Staphylococcus aureus; MurJ; buffer; detergent screening; lipid II Flippase; protein purification |
| المشرفين على المادة: | 0 (Bacterial Proteins) 0 (Detergents) 0 (muramyl-NAc-(pentapeptide)pyrophosphoryl-undecaprenol) 0 (Uridine Diphosphate N-Acetylmuramic Acid) |
| تواريخ الأحداث: | Date Created: 20240705 Date Completed: 20240705 Latest Revision: 20240713 |
| رمز التحديث: | 20240713 |
| DOI: | 10.2174/0109298665316374240531113258 |
| PMID: | 38967080 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1875-5305 |
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| DOI: | 10.2174/0109298665316374240531113258 |