Journal
/
Biochemistry
/
Combining X-Ray Crystallography with Small Angle X-Ray Scattering to Model Unstructured Regions of Nsa1 from S. Cerevisiae
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Combining X-Ray Crystallography with Small Angle X-Ray Scattering to Model Unstructured Regions of Nsa1 from S. Cerevisiae

Combining X-Ray Crystallography with Small Angle X-Ray Scattering to Model Unstructured Regions of Nsa1 from S. Cerevisiae

DOI:
10.3791/56953-v

•

09:15 min

•

January 10, 2018

•

, ,
1Signal Transduction Laboratory, National Institutes of Environmental Health Sciences, Department of Health and Human Services,National Institutes of Health

Chapters

  • 00:05Title
  • 00:47Nsa1 (Nop7 Associated 1) Protein Purification
  • 03:43Proteolytic Screening
  • 05:14Small-Angle X-Ray Scattering (SAXS) Data Processing, Comparison of Concentration Series, and Ensemble Modeling
  • 07:31Results: Modeling the Disordered C-Terminus Region of Nsa1
  • 08:37Conclusion

Summary

Automatic Translation

Automatic Translation

This method describes the cloning, expression, and purification of recombinant Nsa1 for structural determination by X-ray crystallography and small-angle X-ray scattering (SAXS), and is applicable for the hybrid structural analysis of other proteins containing both ordered and disordered domains.

Tags

X-ray CrystallographySmall Angle X-ray Scattering (SAXS)Structural AnalysisProtein StructureRecombinant ProteinUnstructured RegionsNsa1S. CerevisiaeHybrid Structural AnalysisProtein FunctionProtein DynamicsProtein AggregationPurificationAffinity ChromatographyGel FiltrationSDS-PAGETEV ProteaseCentrifugal Filtration

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Characterization of Glycoproteins with the Immunoglobulin Fold by X-Ray Crystallography and Biophysical Techniques

Contrast-Matching Detergent in Small-Angle Neutron Scattering Experiments for Membrane Protein Structural Analysis and Ab Initio Modeling

Structural Studies of Macromolecules in Solution using Small Angle X-Ray Scattering

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization

Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)

An All-in-one Sample Holder for Macromolecular X-ray Crystallography with Minimal Background Scattering

Cell Culture on Silicon Nitride Membranes and Cryopreparation for Synchrotron X-ray Fluorescence Nano-analysis

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050

Fixed Target Serial Data Collection at Diamond Light Source

Motility of Single Molecules and Clusters of Bi-Directional Kinesin-5 Cin8 Purified from S. cerevisiae Cells

Read Article