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Expression and Purification of a Matrix Metalloprotease Transmembrane Domain in Escherichia coli

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Matrix Metalloproteases

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

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Abstract

Membrane tethered matrix metalloproteases are bound to the plasma membrane by a glycosylphosphatidylinositol-anchor or a transmembrane domain. To date, most studies of membrane-bound matrix metalloprotease have focused on the globular catalytic and protein–protein interaction domains of these enzymes. However, the transmembrane domains have been poorly studied even though they are known to mediate intracellular signaling via interaction with various cellular proteins. The expression and purification of the transmembrane domain of these proteins can be challenging due to their hydrophobic nature. In this chapter we describe the purification of a transmembrane domain for a membrane-bound matrix metalloprotease expressed in E. coli and its initial characterization by NMR spectroscopy.

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Authors and Affiliations

  1. Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC, 3052, Australia

    Charles A. Galea

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  1. Charles A. Galea
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Correspondence to Charles A. Galea .

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  1. Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia

    Charles A. Galea

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Galea, C.A. (2017). Expression and Purification of a Matrix Metalloprotease Transmembrane Domain in Escherichia coli . In: Galea, C. (eds) Matrix Metalloproteases. Methods in Molecular Biology, vol 1579. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6863-3_2

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  • DOI: https://doi.org/10.1007/978-1-4939-6863-3_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6861-9

  • Online ISBN: 978-1-4939-6863-3

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