Activity-dependent Interdomain Dynamics of Matrix Metalloprotease-1 on Fibrin

Authors

Kumar, L., Planas-Iglesias, J., Harms, C., Kamboj, S., Wright, D., Klein-Seetharaman, J., Sarkar, S. K.

Source

SCIENTIFIC REPORTS 10: 1-14 (2020)

Abstract

The roles of protein conformational dynamics and allostery in function are well-known. However, the roles that interdomain dynamics have in function are not entirely understood. We used matrix metalloprotease-1 (MMP1) as a model system to study the relationship between interdomain dynamics and activity because MMP1 has diverse substrates. Here we focus on fibrin, the primary component of a blood clot. Water-soluble fibrinogen, following cleavage by thrombin, self-polymerize to form water-insoluble fibrin. We studied the interdomain dynamics of MMP1 on fibrin without crosslinks using single-molecule Forster Resonance Energy Transfer (smFRET). We observed that the distance between the catalytic and hemopexin domains of MMP1 increases or decreases as the MMP1 activity increases or decreases, respectively. We modulated the activity using (1) an active site mutant (E219Q) of MMP1, (2) MMP9, another member of the MMP family that increases the activity of MMP1, and (3) tetracycline, an inhibitor of MMP1. We fitted the histograms of smFRET values to a sum of two Gaussians and the autocorrelations to an exponential and power law. We modeled the dynamics as a two-state Poisson process and calculated the kinetic rates from the histograms and autocorrelations. Activity-dependent interdomain dynamics may enable allosteric control of the MMP1 function.

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Citation

Kumar, L., Planas-Iglesias, J., Harms, C., Kamboj, S., Wright, D., Klein-Seetharaman, J., Sarkar, S. K., 2020: Activity-dependent Interdomain Dynamics of Matrix Metalloprotease-1 on Fibrin. Scientific Reports 10: 1-14.

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