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oai:recercat.cat:20.500.12327/12402025-10-22T11:34:11Zcom_2072_4428com_2072_4427col_2072_487898

00925njm 22002777a 4500 dc Carratalá, Jose Vicente author Gifre-Renom, Laia author Roca-Pinilla, Ramon author Villaverde, Antonio author Alomar, Oscar author Garcia-Fruitos, Elena author Sánchez, Julieta María author Ferrer-Miralles, Neus author 2021-03-16 A detailed workflow to analyze the physicochemical characteristics of mammalian matrix metalloproteinase (MMP-9) protein species obtained from protein aggregates (inclusion bodies—IBs) was followed. MMP-9 was recombinantly produced in the prokaryotic microbial cell factories Clearcoli (an engineered form of Escherichia coli) and Lactococcus lactis, mainly forming part of IBs and partially recovered under non-denaturing conditions. After the purification by affinity chromatography of solubilized MMP-9, four protein peaks were obtained. However, so far, the different conformational protein species forming part of IBs have not been isolated and characterized. Therefore, with the aim to link the physicochemical characteristics of the isolated peaks with their biological activity, we set up a methodological approach that included dynamic light scattering (DLS), circular dichroism (CD), and spectrofluorometric analysis confirming the separation of subpopulations of conformers with specific characteristics. In protein purification procedures, the detailed analysis of the individual physicochemical properties and the biological activity of protein peaks separated by chromatographic techniques is a reliable source of information to select the best-fitted protein populations. Carratalá, Jose Vicente, Laia Gifre-Renom, Ramon Roca-Pinilla, Antonio Villaverde, Anna Arís, Elena Garcia-Fruitós, Julieta María Sánchez, and Neus Ferrer-Miralles. 2021. "Selecting Subpopulations Of High-Quality Protein Conformers Among Conformational Mixtures Of Recombinant Bovine MMP-9 Solubilized From Inclusion Bodies". International Journal Of Molecular Sciences 22 (6): 3020. doi:10.3390/ijms22063020. 1661-6596 http://hdl.handle.net/20.500.12327/1240 https://doi.org/10.3390/ijms22063020 Selecting Subpopulations of High-Quality Protein Conformers among Conformational Mixtures of Recombinant Bovine MMP-9 Solubilized from Inclusion Bodies