دورية أكاديمية
Expression, purification, and characterization of anuran saxiphilins using thermofluor, fluorescence polarization, and isothermal titration calorimetry.
| العنوان: | Expression, purification, and characterization of anuran saxiphilins using thermofluor, fluorescence polarization, and isothermal titration calorimetry. |
|---|---|
| المؤلفون: | Chen Z; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94158-2330, USA., Zakrzewska S; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94158-2330, USA., Hajare HS; Department of Chemistry, Stanford University, Stanford, CA 94305, USA., Du Bois J; Department of Chemistry, Stanford University, Stanford, CA 94305, USA. Electronic address: jdubois@stanford.edu., Minor DL Jr; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94158-2330, USA; Departments of Biochemistry and Biophysics, and Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158-2330, USA; California Institute for Quantitative Biomedical Research, University of California, San Francisco, San Francisco, CA 94158-2330, USA; Kavli Institute for Fundamental Neuroscience, University of California, San Francisco, San Francisco, CA 94158-2330, USA; Molecular Biophysics and Integrated Bio-imaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA. Electronic address: daniel.minor@ucsf.edu. |
| المصدر: | STAR protocols [STAR Protoc] 2024 Mar 15; Vol. 5 (1), pp. 102792. Date of Electronic Publication: 2023 Dec 20. |
| 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: | Saxitoxin*/metabolism , Neurotoxins*, Calorimetry ; Fluorescence Polarization |
| مستخلص: | Anuran saxiphilins (Sxphs) are "toxin sponge" proteins thought to prevent the lethal effects of small-molecule neurotoxins through sequestration. Here, we present a protocol for the expression, purification, and characterization of Sxphs. We describe steps for using thermofluor, fluorescence polarization, and isothermal titration calorimetry assays that probe Sxph:saxitoxin interactions using a range of sample quantities. These assays are generalizable and can be used for other paralytic shellfish poisoning toxin-binding proteins. For complete details on the use and execution of this protocol, please refer to Chen et al. (2022). 1 . Competing Interests: Declaration of interests J.D.B. is a cofounder and holds equity shares in SiteOne Therapeutics, Inc., a start-up company interested in developing subtype-selective modulators of sodium channels. Patent applications have been filed for this work. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.) |
| معلومات مُعتمدة: | R01 GM117263 United States GM NIGMS NIH HHS |
| فهرسة مساهمة: | Keywords: Protein Biochemistry; Protein expression and purification; Structural Biology |
| المشرفين على المادة: | 35523-89-8 (Saxitoxin) 0 (Neurotoxins) |
| تواريخ الأحداث: | Date Created: 20231222 Date Completed: 20240320 Latest Revision: 20240320 |
| رمز التحديث: | 20240320 |
| مُعرف محوري في PubMed: | PMC10776646 |
| DOI: | 10.1016/j.xpro.2023.102792 |
| PMID: | 38133955 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2666-1667 |
|---|---|
| DOI: | 10.1016/j.xpro.2023.102792 |