Thermal stability of human matrix metalloproteinases

Noemi Meraz-Cruz; Felipe Vadillo-Ortega; Aura M Jiménez-Garduño; Alicia Ortega

doi:10.1016/j.heliyon.2020.e03865

Thermal stability of human matrix metalloproteinases

Heliyon. 2020 May 3;6(5):e03865. doi: 10.1016/j.heliyon.2020.e03865. eCollection 2020 May.

Authors

Noemi Meraz-Cruz^{1

2}, Felipe Vadillo-Ortega^{1

2}, Aura M Jiménez-Garduño^{1

3}, Alicia Ortega¹

Affiliations

¹ Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, 04510, Mexico City, Mexico.
² UNAM School of Medicine Branch, National Institute of Genomic Medicine, Mexico.
³ Health Sciences Department, School of Sciences, Universidad de Las Americas Puebla, San Andrés Cholula Puebla CP72810, Puebla, Mexico.

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.

Keywords: Biochemistry; Biological sciences; Chemistry; Collagenase; Differential scanning calorimetry; Matrilysin; Metalloproteinases; Protein denaturation; Proteins; Stromelysin.