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ZIP Metal Ion Transporters
- 1st Edition, Volume 687 - August 31, 2023
- Editor: Jian Hu
- Language: English
- Hardback ISBN:9 7 8 - 0 - 3 2 3 - 9 5 7 7 6 - 2
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 9 5 7 7 7 - 9
ZIP Metal Ion Transporters, Volume 684 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a va… Read more
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Request a sales quoteZIP Metal Ion Transporters, Volume 684 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a variety of topics, including Biochemistry, Biophysics and Structural Biology, Structure determination of a bacterial ZIP in lipidic cubic phase, Cell-based metal transport assay for animal and plant ZIPs, Measurement of metal binding affinity and transport activity in ZIP transporters using spectroscopic methods, Considerations in production of the (prokaryotic) ZIP family transporters for structural and functional studies, Metal-protein interactions investigated using model systems: thermodynamic, spectroscopic and computational methods, and more.
Additional chapters cover Getting Zinc Into and Out of Cells, In-silico modeling of ZIP9 (TBD), Cellular Function and Regulation, Methods to visualize zinc transporter proteins of the SLC39A family in cells, Assessing metal ion transporting activity of ZIPs: intracellular zinc and iron detection, In vitro studies for studying manganese transport and homeostasis, and much more.
- Provides the authority and expertise of leading contributors from an international board of authors
- Presents the latest release in Methods in Enzymology serials
- Updated release includes the latest information on ZIP Metal Ion Transporters
- Cover image
- Title page
- Table of Contents
- Methods in ENZYMOLOGY
- METHODS IN ENZYMOLOGY
- Methods in ENZYMOLOGY
- Copyright
- Contributors
- Preface
- Chapter One: Considerations in production of the prokaryotic ZIP family transporters for structural and functional studies
- Abstract
- 1 Introduction
- 2 Plasmids construction
- 3 Expression screens
- 4 Detergents screens
- 5 Target protein purifications
- 6 Stability screens
- 7 Function assays
- 8 Summary
- Acknowledgements
- References
- Chapter Two: Expression, purification, crystallization of a ZIP metal transporter from Bordetella bronchiseptica (BbZIP)
- Abstract
- 1 Introduction
- 2 Overexpression of BbZIP
- 3 Purification of BbZIP
- 4 Crystallization of BbZIP
- 5 Summary
- Acknowledgments
- References
- Chapter Three: Expression, purification, and crystallization of the extracellular domain of a mammalian ZIP4
- Abstract
- 1 Introduction
- 2 Before you begin
- 3 Key resource table
- 4 Materials and equipment
- 5 Step-by-step method details
- 6 Expected outcomes
- 7 Discussion
- References
- Chapter Four: Methods to visualise zinc transporter proteins of the SLC39A family in cells
- Highlights
- Abstract
- 1 Introduction
- 2 General methods
- 3 Special treatment considerations and probe choice
- 4 Transfection
- 5 Western blotting
- 6 Immunofluorescence
- 7 Proximity ligation assay
- 8 Summary and conclusions
- References
- Chapter Five: A simple and highly reproducible method for the detection of erythrocyte membrane ZIP metal transporters by immunoblotting
- Abstract
- 1 Introduction
- 2 Isolation of erythrocyte membrane from mouse blood
- 3 Detection of ZIP proteins by immunoblotting method
- 4 Discussion
- Institutional review board statement
- References
- Chapter Six: Single-molecule quantification of the oligomeric state of ZIP transporters in mammalian cells with fluorescence correlation spectroscopy
- Abstract
- 1 Introduction
- 2 General safety measures
- 3 Molecular design of fluorescent protein tagged gene fusions
- 4 Mammalian cell culture
- 5 Confocal microscopy set up
- 6 Fluorescence correlation spectroscopy data analysis
- 7 Summary and conclusions
- Acknowledgments
- References
- Chapter Seven: Cell-based transport assay to study kinetics and substrate specificity of human ZIPs
- Abstract
- 1 Introduction
- 2 Determination of zinc ion transport kinetics of human ZIP4 using cell-based transport assay
- 3 Comparison of substrate specificities of human ZIP8 and human ZIP4 using cell-based transport assay
- 4 Summary
- Acknowledgments
- References
- Chapter Eight: Assessing metal ion transporting activity of ZIPs: Intracellular zinc and iron detection
- Abstract
- 1 Introduction
- 2 Intracellular zinc detection
- 3 Intracellular iron detection
- 4 Summary
- Acknowledgments
- References
- Chapter Nine: In vitro studies of manganese transport and homeostasis
- Abstract
- 1 Introduction
- 2 General method and statistical analysis
- 3 Vector construction and site-directed mutagenesis of SLC39A8
- 4 Transient transfection of SLC39A8 and its mutants into HeLa cells
- 5 Measurement of total Mn concentration
- 6 54Mn uptake assay
- 7 GPP130 degradation assay
- 8 Immunofluorescence imaging
- 9 Summary and conclusions
- Acknowledgments
- References
- Chapter Ten: Simple in vitro method to evaluate ZIP zinc transport ability through zinc transporter 1 and metallothionein expression measurements
- Abstract
- 1 Introduction
- 2 Evaluation of Zn transport of transfected ZIPs by analyzing ZNT1 expression using immunofluorescence staining
- 3 Evaluation of Zn transport of ZIPs by analyzing ZNT1 expression with immunoblot analysis
- 4 Use of ZNT1 expression to monitor the decreased Zn levels mediated by loss of ZIP expression
- 5 Monitoring ZIPs-driven ZNT1 expression in polarized cells
- 6 Evaluation of the Zn transport ability of ZIPs through analyzing MT expression
- 7 In vitro screening through evaluating dietary components enhancing ZIP4-mediated Zn uptake: Application of the ZIP4-mediated MT expression response
- 8 Confirmation of ZIP4 induction by the food components in the in vitro screening system
- 9 Advantages
- 10 Limitations
- 11 Conclusion
- Acknowledgments
- References
- Chapter Eleven: Internalization and trafficking of zinc transporters
- Abstract
- 1 Introduction
- 2 Western blot
- 3 Immunofluorescence and confocal microscopy
- 4 Conclusion and future perspectives
- References
- Chapter Twelve: Getting zinc into and out of cells
- Abstract
- 1 Introduction
- 2 Computational details
- 3 Proposed mechanism of YiiP transporters
- 4 Proposed mechanism of ZIP transporters
- 5 Conclusions
- Acknowledgements
- References
- Chapter Thirteen: Metal–protein solution interactions investigated using model systems: Thermodynamic and spectroscopic methods
- Abstract
- 1 Introduction
- 2 Instrumental techniques
- 3 A case study: The periplasmic protein ZinT
- 4 A case study: The ZIP protein Zrt2
- 5 Final remarks
- Acknowledgements
- References
- No. of pages: 412
- Language: English
- Edition: 1
- Volume: 687
- Published: August 31, 2023
- Imprint: Academic Press
- Hardback ISBN: 9780323957762
- eBook ISBN: 9780323957779
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