title: Expression and Purification of a Matrix Metalloprotease Transmembrane Domain in Escherichia coli.
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.
authors.affiliation: Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC, 3052, Australia. charles.galea@monash.edu.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Galea
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development*; D020782:Matrix Metalloproteinases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D019906:Nuclear Magnetic Resonance, Biomolecular; D000072417:Protein Domains; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D020782:Matrix Metalloproteinases
keywords: Isotopically labeled; Matrix metalloproteases; Membrane-anchored MMP; NMR spectroscopy; Protein expression; Transmembrane domain