Expression of recombinant matrix metalloproteinases in Escherichia coli.
Matrix metalloproteinases (MMPs) are a group of zinc-dependent endopeptidases that are capable of cleaving all of the components of the extracellular matrix (ECM). The role that the MMPs play in normal and pathological conditions has long been of interest. The mechanisms by which the MMPs cleave the different components of the ECM have been examined extensively. Some of these studies have been made possible, in part, by the ability to express recombinant MMPs. These recombinant MMPs have been utilized in both structural and functional studies. In addition, future studies can benefit from the availability of recombinant MMPs. Recombinant MMPs have been expressed in mammalian and bacterial recombinant expression systems. The most common bacterial expression system employed for this has been the utilization of expression plasmids in Escherichia coli. This has resulted in the production of a large amount of protein in a short period of time. The expression of a recombinant truncated form of human stromelysin-1 (MMP-3) will be used to illustrate the methods utilized for the expression of a MMP in E. coli. This will include discussions about the expression vector, the cloning of the MMP cDNA into the expression vector, protein induction, protein extraction, protein refolding and purification, and protein characterization.
Department of Oral Biology, Indiana University School of Dentistry, Indianapolis, IN, USA.
L Jack
LJ
Windsor
Darin L
DL
Steele
D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004789:Enzyme Activation; D004790:Enzyme Induction; D004926:Escherichia coli / Q000378:metabolism*; D006801:Humans; D020782:Matrix Metalloproteinases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism*
D011994:Recombinant Proteins; D020782:Matrix Metalloproteinases