Following matrix metalloproteinases activity near the cell boundary by infrared micro-spectroscopy.
Matrix Metalloproteinases (MMPs) are cell-secreted soluble and membrane-tethered enzymes that degrade extracellular matrix (ECM) proteins. These proteases play a key role in diverse physiological and pathological processes, including embryonic development, wound repair, inflammatory diseases and cancer. Yet, there is insufficient knowledge on the mode by which cell-produced MMPs conduct their action on the ECM. Specifically, the localization and the mode of the degradation within the pericellular space are of great interest. To provide new insights to these questions we utilized Fourier transform infrared (FTIR) micro-spectroscopy to follow proteolytic processes, induced by invasive cancer cells, on insoluble collagen-based matrices. Here we show that FTIR micro-spectroscopy have a great potential for monitoring degradation events near cells. Using this tool we demonstrate that the net proteolysis is unevenly distributed around the cell boundary. The degradation patterns show different levels of proteolytic activity by MMPs within the pericellular space. In addition, our spectral analysis suggests that the enzymatic proteolysis of the collagen-based matrices induces unwinding of the triple helical structures of the macromolecules within the collagen network.
Federman
Silvina
S
Department of Structural Biology, The Weizmann Institute of Science, 76100, Rehovot, Israel.
Miller
Lisa M
LM
Sagi
Irit
I
D003094:Collagen; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000378:metabolism*; D005354:Fibrosarcoma / Q000378:metabolism* / Q000473:pathology; D005780:Gelatin; D006801:Humans; D007797:Laminin; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D011509:Proteoglycans; D017550:Spectroscopy, Fourier Transform Infrared; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D005780:Gelatin; D003094:Collagen; D020782:Matrix Metalloproteinases