From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins

13.4K Views

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09:55 min

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July 4th, 2016

DOI :

10.3791/53245-v

July 4th, 2016

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Nicholas Noinaj¹, Stephen Mayclin², Ann M. Stanley²^,³, Christine C. Jao²^,³, Susan K. Buchanan²

¹Department of Biological Sciences, Markey Center for Structural Biology, Purdue University, ²National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health, ³National Institute of General Medical Sciences (NIGMS), National Institutes of Health

Summary

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Keywords Outer Membrane Proteins

Beta barrel Proteins

Membrane Protein Folding

Protein Expression

Protein Purification

Protein Crystallization

Chapters in this video

0:05

Title

0:46

In Vivo Membrane-targeted Expression

3:09

Isolation from Membrane Fractions

4:41

Purification Using Immobilized Metal-affinity Chromatography (IMAC), Ion-exchange, and Gel Filtration

7:09

Heat Modifiability Assay

8:35

Results: Crystals to Structure Determination for the Putative Vaccine Target YiuR from Yersinia pestis

9:29

Conclusion

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