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Imaging Intermediate Filaments and Microtubules with 2-dimensional Direct Stochastic Optical Reconstruction Microscopy
Published on: March 6, 2018
Characterizing the function-related specific assembly pattern of matrix metalloproteinase-14 by dSTORM imaging
Dian Ge1, Junling Chen1, Zhiyong Zhao1
1Improve-WUST Joint Laboratory of Advanced Technology for Point-of-Care Testing and Precision Medicine, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, 947 Heping Street, Wuhan, Hubei, 430081, China.
Abstract:
As transmembrane proteolytic enzyme, matrix metalloproteinase-14 (MMP14) regulates cell migration and cancer metastasis, but how it works at the single molecule level is unclear. Molecular localization is closely related to its function, and revealing its spatial assemble details is thus helpful to understand bio-function. Here, we apply aptamer probe and dSTORM to characterize MMP14 distribution. With demonstrating labeling properties of the probe, we investigate the specific distributed pattern of MMP14 on various cell membranes with different migratory capacities, and find that MMP14 mostly aggregate in clustering state, which becomes more significant with enhancing its hydrolysis efficiency on high-migratory cells. Lots of MMP14 are revealed to be co-localized with its substrate PTK7, and this colocalization decreases with weakening cell migration, suggesting that MMP14 may coordinate cell migration by altering its spatial relationship with substrate proteins. This work will promote a deep understanding of the roles of MMP14 in cell migration and cancer metastasis.
Insights
Matrix metalloproteinase-14 (MMP14) aggregates on cell membranes, coordinating cell migration. Its clustering increases with hydrolysis efficiency and co-localization with substrate PTK7 on highly migratory cells.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Matrix metalloproteinase-14 (MMP14) is a transmembrane enzyme regulating cell migration and cancer metastasis.
- Understanding MMP14's single-molecule function requires detailed knowledge of its spatial organization.
Purpose of the Study:
- To characterize the single-molecule distribution and spatial assembly of MMP14 on cell membranes.
- To investigate the relationship between MMP14 localization, migratory capacity, and substrate interaction.
Main Methods:
- Development and application of aptamer probes for MMP14 labeling.
- Utilizing direct Stochastic Optical Reconstruction Microscopy (dSTORM) for high-resolution imaging.
- Analyzing MMP14 co-localization with its substrate PTK7.
Main Results:
- MMP14 predominantly forms clusters on cell membranes.
- MMP14 clustering is more pronounced in high-migratory cells and correlates with enhanced hydrolysis efficiency.
- Significant co-localization of MMP14 with PTK7 was observed, decreasing as cell migration weakens.
Conclusions:
- MMP14 spatial organization, particularly its clustering and co-localization with PTK7, is crucial for coordinating cell migration.
- This study provides insights into MMP14's role in cell migration and cancer metastasis at the single-molecule level.
