Purification of Native Dentilisin Complex from Treponema denticola by Preparative Continuous Polyacrylamide Gel Electrophoresis and Functional Analysis by Gelatin Zymography
Pachiyappan Kamarajan,
John Timm,
M. Paula Goetting-Minesky,
Erin Malone,
Sean Ganther,
Allan Radaic,
Christian Tafolla,
J. Fenno,
Yvonne Kapila
Affiliations
Pachiyappan Kamarajan
Department of Biosystems and Function, School of Dentistry, University of California Los Angeles, Los Angeles, CA, USA
John Timm
Department of Biologic and Materials Sciences & Prosthodontics, School of Dentistry, University of Michigan, Ann Arbor, MI, USA
M. Paula Goetting-Minesky
Department of Biologic and Materials Sciences & Prosthodontics, School of Dentistry, University of Michigan, Ann Arbor, MI, USA
Erin Malone
Department of Orofacial Sciences, School of Dentistry, University of California San Francisco, San Francisco, CA, USA
Sean Ganther
Department of Orofacial Sciences, School of Dentistry, University of California San Francisco, San Francisco, CA, USA
Allan Radaic
Department of Biosystems and Function, School of Dentistry, University of California Los Angeles, Los Angeles, CA, USA
Christian Tafolla
Department of Orofacial Sciences, School of Dentistry, University of California San Francisco, San Francisco, CA, USA
J. Fenno
Department of Biologic and Materials Sciences & Prosthodontics, School of Dentistry, University of Michigan, Ann Arbor, MI, USA
Yvonne Kapila
Department of Biosystems and Function, School of Dentistry, University of California Los Angeles, Los Angeles, CA, USADepartment of Orofacial Sciences, School of Dentistry, University of California Los Angeles, Los Angeles, CA, USA
Periodontal disease is characterized by the destruction of the hard and soft tissues comprising the periodontium. This destruction translates to a degradation of the extracellular matrices (ECM), mediated by bacterial proteases, host-derived matrix metalloproteinases (MMPs), and other proteases released by host tissues and immune cells. Bacterial pathogens interact with host tissue, triggering adverse cellular functions, including a heightened immune response, tissue destruction, and tissue migration. The oral spirochete Treponema denticola is highly associated with periodontal disease. Dentilisin, a T. denticola outer membrane protein complex, contributes to the chronic activation of pro-MMP-2 in periodontal ligament (PDL) cells and triggers increased expression levels of activators and effectors of active MMP-2 in PDL cells. Despite these advances, no mechanism for dentilisin-induced MMP-2 activation or PDL cytopathic behaviors leading to disease is known. Here, we describe a method for purification of large amounts of the dentilisin protease complex from T. denticola and demonstrate its ability to activate MMP-2, a key regulator of periodontal tissue homeostasis. The T. denticola dentilisin and MMP-2 activation model presented here may provide new insights into the dentilisin protein and identify potential therapeutic targets for further research.
