Expression and purification of a matrix metalloprotease transmembrane domain in Escherichia coli

Charles A Galea

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

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.

Original languageEnglish
Title of host publicationMatrix Metalloproteases
Subtitle of host publicationMethods and Protocols
EditorsCharles A Galea
PublisherSpringer
Pages17-33
Number of pages17
Volume1579
ISBN (Electronic)9781493968633
ISBN (Print)9781493968619
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1579
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Isotopically labeled
  • Matrix metalloproteases
  • Membrane-anchored MMP
  • NMR spectroscopy
  • Protein expression
  • Transmembrane domain

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