Abstract
Due to their central roles in tumor growth and invasion, milligram-level amounts of active MMPs are frequently required for cancer research and development of chemical or biological MMP inhibitors. Here we describe methods for functional production of catalytic domains of MMPs (cdMMPs) in E. coli periplasm without refolding or activation process. We demonstrate applications of this straightforward approach for cdMMP-9, cdMMP-14, and cdMMP-14 mutants.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Sela-Passwell N, Rosenblum G, Shoham T et al (2010) Structural and functional bases for allosteric control of MMP activities: can it pave the path for selective inhibition? Biochim Biophys Acta 1803(1):29–38
Jiang A, Lehti K, Wang X et al (2001) Regulation of membrane-type matrix metalloproteinase 1 activity by dynamin-mediated endocytosis. Proc Natl Acad Sci U S A 98(24):13693–13698
Tetu B, Brisson J, Wang CS et al (2006) The influence of MMP-14, TIMP-2 and MMP-2 expression on breast cancer prognosis. Breast Cancer Res 8(3):R28
Mimori K, Ueo H, Shirasaka C et al (2001) Clinical significance of MT1-MMP mRNA expression in breast cancer. Oncol Rep 8(2):401–403
Genís L, Gálvez BG, Gonzalo P et al (2006) MT1-MMP: universal or particular player in angiogenesis? Cancer Metastasis Rev 25(1):77–86
Morrison CJ, Butler GS, Rodríguez D et al (2009) Matrix metalloproteinase proteomics: substrates, targets, and therapy. Curr Opin Cell Biol 21(5):645–653
Devy L, Dransfield DT (2011) New Strategies for the next generation of matrix-metalloproteinase inhibitors: selectively targeting membrane-anchored MMPs with therapeutic antibodies. Biochem Res Int 2011:191670
Koo HM, Kim JH, Hwang IK et al (2002) Refolding of the catalytic and hinge domains of human MT1-mMP expressed in Escherichia coli and its characterization. Mol Cells 13(1):118–124
Lichte A, Kolkenbrock H, Tschesche H (1996) The recombinant catalytic domain of membrane-type matrix metalloproteinase-1 (MT1-MMP) induces activation of progelatinase A and progelatinase A complexed with TIMP-2. FEBS Lett 397(2–3):277–282
Baneyx F, Mujacic M (2004) Recombinant protein folding and misfolding in Escherichia coli. Nat Biotechnol 22(11):1399–1408
Will H, Atkinson SJ, Butler GS et al (1996) The soluble catalytic domain of membrane type 1 matrix metalloproteinase cleaves the propeptide of progelatinase A and initiates autoproteolytic activation. Regulation by TIMP-2 and TIMP-3. J Biol Chem 271(29):17119–17123
Knäuper V, Will H, López-Otin C et al (1996) Cellular mechanisms for human procollagenase-3 (MMP-13) activation. Evidence that MT1-MMP (MMP-14) and gelatinase a (MMP-2) are able to generate active enzyme. J Biol Chem 271(29):17124–17131
Nam DH, Ge X (2016) Direct production of functional matrix metalloproteinase-14 without refolding or activation and its application for in vitro inhibition assays. Biotechnol Bioeng 113(4):717–723
Hayhurst A, Happe S, Mabry R et al (2003) Isolation and expression of recombinant antibody fragments to the biological warfare pathogen Brucella melitensis. J Immunol Methods 276(1–2):185–196
Nam DH, Ge X (2013) Development of periplasmic FRET-based screening method for protease inhibitory antibodies. Biotechnol Bioeng 110(11):2856–2864
Nam DH, Rodriguez C, Remacle AG et al (2016) Active-site MMP-selective antibody inhibitors discovered from convex paratope synthetic libraries. Proc Natl Acad Sci U S A 113(52):14970–14975
Lee KB, Nam DH, Nuhn J et al (2017) Direct expression of active human tissue inhibitors of Metalloproteinase by periplasmic secretion in Escherichia coli. Microb Cell Factories 16:73
Goldman ER, Hayhurst A, Lingerfelt BM et al (2003) 2,4,6-Trinitrotoluene detection using recombinant antibodies. J Environ Monit 5(3):380–383
Acknowledgments
This work was supported by National Science Foundation the Faculty Early Career Development (CAREER) Program 1453645 and National Institutes of Health Grant R01 GM115672 to X.G.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Nam, D.H., Lee, K.B., Ge, X. (2018). Functional Production of Catalytic Domains of Human MMPs in Escherichia coli Periplasm. In: Cal, S., Obaya, A. (eds) Proteases and Cancer. Methods in Molecular Biology, vol 1731. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7595-2_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7595-2_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7594-5
Online ISBN: 978-1-4939-7595-2
eBook Packages: Springer Protocols