Recombinant human 92-kDa type IV collagenase/gelatinase from baculovirus-infected insect cells: expression, purification, and characterization

Protein Expression and Purification
H J GeorgeR A Copeland

Abstract

Human 92-kDa type IV collagenase/gelatinase (MMP9) has been expressed in insect cells and secreted into the cell medium via a baculovirus expression system. The expression level of the proenzyme from Trichoplusia ni cells was estimated to be > = 300 mg/L of cell medium. The recombinant protein was purified in a single step using heparin-affinity chromatography with an overall yield of ca. 70%. The purified zymogen could be activated in vitro using 4-aminophenylmercuric acetate to yield an active protease. Kinetic analysis of the activated recombinant enzyme demonstrates that this material is comparable to activated MMP9 from natural human sources. The recombinant enzyme provides a useful source of protein for a variety of biochemical and biophysical studies aimed at elucidating the structure and function of human MMP9.

References

Jan 1, 1995·Methods in Enzymology·N D Rawlings, A J Barrett
Jan 1, 1995·Methods in Enzymology·G Murphy, T Crabbe

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Citations

Aug 22, 2003·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·A TongE Monsonego-Ornan
Jun 22, 2005·Trends in Biotechnology·K M Helena NevalainenPeter L Bergquist
Jul 27, 1999·Protein Expression and Purification·N RoyG Das
Feb 24, 2015·Matrix Biology : Journal of the International Society for Matrix Biology·Kazuhiro YamamotoLinda Troeberg
Oct 14, 2004·Applied Microbiology and Biotechnology·F R Schmidt
Jan 20, 2006·The FEBS Journal·Lillian ZernichowSvein O Kolset
Mar 9, 2013·PloS One·Kotakonda ArunasriSisinthy Shivaji

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