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Bacterial Expression and Purification of Human Matrix Metalloproteinase-3 using Affinity Chromatography
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Biochemistry
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JoVE Journal Biochemistry
Bacterial Expression and Purification of Human Matrix Metalloproteinase-3 using Affinity Chromatography

Bacterial Expression and Purification of Human Matrix Metalloproteinase-3 using Affinity Chromatography

DOI:
10.3791/63263-v

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07:32 min

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March 30, 2022

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, ,
1Department of Chemical & Materials Engineering,University of Nevada, Reno

Chapters

  • 00:04Introduction
  • 00:52MMP Expression
  • 03:46MMP Purification
  • 04:27MMP Refolding
  • 05:12MMP Activation
  • 05:46Results: Purification and Activation of Hisx6-Pro-MMP-3cd From BL21(DE3) and R2DP Cells
  • 06:38Conclusion

Summary

Automatic Translation

Automatic Translation

His-tag purification, dialysis, and activation are employed to increase yields of soluble, active matrix metalloproteinase-3 catalytic domain protein expression in bacteria. Protein fractions are analyzed via SDS-PAGE gels.

Tags

Bacterial ExpressionRecombinant MMP-3Affinity ChromatographyE. ColiR2DP Cell StrainMMP InhibitorsProtein PurificationProtein FoldingPost-translational ProcessingIPTG InductionCell LysisInclusion Body SolubilizationSonicationCentrifugation

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