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Bioquímica

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

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Bioquímica

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JoVE Journal Bioquímica

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

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

DOI:

10.3791/57901-v

•

10:27 min

•

October 21, 2018

•

Ryan C. Oliver, Swe-Htet Naing, Kevin L. Weiss, Sai Venkatesh Pingali, Raquel L. Lieberman, Volker S. Urban

¹Neutron Scattering Division,Oak Ridge National Laboratory, ²School of Chemistry and Biochemistry,Georgia Institute of Technology

Capítulos

00:04Título
00:54Determine Neutron Contrast Match Points and Necessary Contrast for Protein Measurement
02:05Express and Purify the Membrane Protein of Interest
03:25Collect SANS Data and Reduce SANS Data from 2D Image to 1D Plot
04:48Analyze Data for Structural Parameters of the Scattering Particle
07:06Create Ab Initio Models from the SANS Data
08:17Results: Visualization of the Experimental SANS Envelope and Overlay with High-resolution Model using PyMOL
09:40Conclusion

Summary

Tadução automática

This protocol demonstrates how to obtain a low-resolution ab initio model and structural details of a detergent-solubilized membrane protein in solution using small-angle neutron scattering with contrast-matching of the detergent.

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