Structural Studies of Matrix Metalloproteinase by X-Ray Diffraction

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1579)

Abstract

Matrix Metalloproteinases (MMPs) are a family of proteolytic enzymes whose endopeptidase activity is dependent on the presence of specific metal ions. MT1-MMP (or MMP-14), which has been implicated in tumor progression and cellular invasion, contains a membrane-spanning region located C-terminal to a hemopexin-like domain and an N-terminal catalytic domain. We recombinantly expressed the catalytic domain of human MT1-MMP in E. coli and purified it from inclusion bodies using a refolding protocol that yielded significant quantities of active protein. Crystals of MT1-MMP were obtained using the vapour diffusion method. Here, we describe the protocols used for crystallization and the data analysis together with the resulting diffraction pattern.

Key words

Metalloproteinase MMP-14 MT1-MMP Crystallization Diffraction 
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Notes

Acknowledgment

We thank Dr. Moran Grossman and Prof. Irit Sagi for providing the plasmid containing human MT1-MMP and Yvonne Brandenburger and Ingeborg Heise for technical assistance. This work was supported by the Cluster of Excellence RESOLV (grant No. EXC 1069) funded by the Deutsche Forschungsgemeinschaft and the Max Planck Society.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Elena Decaneto
    • 1
  • Wolfgang Lubitz
    • 1
  • Hideaki Ogata
    • 1
  1. 1.Max Planck Institute for Chemical Energy ConversionMülheim an der RuhrGermany

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