Thermal stability of human matrix metalloproteinases

Abstract

Matrix metalloproteinases (MMP) are key players in the remodelling of the extracellular matrix under physiological and pathological conditions. Thermodynamic parameters of human recombinant metalloproteinases of the active (rMMP2, 3, 7, 8 and 9) and latent (rPro-MMP2, 3 and 9) forms were obtained by differential scanning calorimetry (DSC). Temperature by itself does not result in autocatalysis of recombinant MMP. The transitions observed by DSC correspond to structural domains of the monomeric protein. In this study, we show the domain organization of these proteins, where the thermal transition (Tm) of the main component is observed at 71.3 C (ProMMP-2); 74.8 C (ProMMP-8); 80.0 C (ProMMP-3); 92.6 C (ProMMP-9) and 98.3 C (ProMMP-7). For MMP-3, this main Tm is related to the catalytic domain (CD). The isolated recombinant CD of MMP-3 unfolds as a single transition at Tm 83.4 C, matching the more stable domain observed in the full-length active form of rMMP-3. The denaturation profile of rProMMP-3 shows the main transition at Tm 80 C, a less stable domain before the propeptide domain (PD) cleavage. Our results indicate that the structural stability of MMP and particularly their CD are not substantially altered after cleavage of the PD. We propose that the thermodynamic parameters obtained by DSC are relevant for the functional study of MMP, particularly to reveal their contribution in complex biological samples in health and disease.

Effect of Different Crosslinkers on Denatured Dentin Collagen’s Biostability, MMP Inhibition and Mechanical Properties

“…Endogenous MMP activity was reduced in both DD samples compared with natural dentin collagen ( p < 0.001). This can be explained by the probable denaturation of the MMP enzymes themselves, hence affecting their ability to cleave collagen [ 43 ]. Another explanation could be the alterations of the specific cleavage sites on the collagen molecule itself, such as Gly-Ile in α1 chain and Gly-leu in α2 [ 44 ] upon which the enzymes act.…”

Contrast enhanced photoacoustic detection of fibrillar collagen in the near infrared region-I

“…1B ). Notably, this Tm value closely resembles that of active enzyme zymogens such as MMP-2 (71.3 °C) and MMP-8 (74.8 °C), 31 indicating a high level of protein stability for the MMP-1 decoy.…”

Paracrine effects of the senescence-associated secretory phenotype decrease cancer cell adhesion

“…Matrix metalloproteinases (MMPs) are a known part of the SASP (Coppe et al 2010). Although many of the MMPs are not expressed in Ovcar8s, and we did not observe major differences in MMP gene expression in cisplatin-induced senescence HGSOC cells ( Table S2 ), we controlled for potential increased MMP secretion through heat inactivated CM by boiling for 20 minutes at temperatures above MMPs melting points (Meraz-Cruz et al 2020). Interestingly, we observed similar phenotypes in heat inactivated SCM ( Fig.…”