@article {ijms23137264,
	title = {Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation},
	journal = {International Journal of Molecular Sciences},
	volume = {23},
	number = {13},
	year = {2022},
	note = {n/a},
	abstract = {<p>Membrane-bound catechol-O-methyltransferase (MBCOMT), present in the brain and involved in the main pathway of the catechol neurotransmitter deactivation, is linked to several types of human dementia, which are relevant pharmacological targets for new potent and nontoxic inhibitors that have been developed, particularly for Parkinson\&rsquo;s disease treatment. However, the inexistence of an MBCOMT 3D-structure presents a blockage in new drugs\&rsquo; design and clinical studies due to its instability. The enzyme has a clear tendency to lose its biological activity in a short period of time. To avoid the enzyme sequestering into a non-native state during the downstream processing, a multi-component buffer plays a major role, with the addition of additives such as cysteine, glycerol, and trehalose showing promising results towards minimizing hMBCOMT damage and enhancing its stability. In addition, ionic liquids, due to their virtually unlimited choices for cation/anion paring, are potential protein stabilizers for the process and storage buffers. Screening experiments were designed to evaluate the effect of distinct cation/anion ILs interaction in hMBCOMT enzymatic activity. The ionic liquids: choline glutamate [Ch][Glu], choline dihydrogen phosphate ([Ch][DHP]), choline chloride ([Ch]Cl), 1- dodecyl-3-methylimidazolium chloride ([C12mim]Cl), and 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) were supplemented to hMBCOMT lysates in a concentration from 5 to 500 mM. A major potential stabilizing effect was obtained using [Ch][DHP] (10 and 50 mM). From the DoE 146\% of hMBCOMT activity recovery was obtained with [Ch][DHP] optimal conditions (7.5 mM) at \&minus;80 \&deg;C during 32.4 h. These results are of crucial importance for further drug development once the enzyme can be stabilized for longer periods of time.</p>
},
	issn = {1422-0067},
	doi = {10.3390/ijms23137264},
	url = {https://www.mdpi.com/1422-0067/23/13/7264},
	author = {Gon{\c c}alves, Ana M. and Sousa, {\^A}ngela and Pedro, Augusto Q. and Rom{\~a}o, Maria J. and Jo{\~a}o A. Queiroz and Gallardo, Eug{\'e}nia and Passarinha, Lu{\'\i}s A.}
}
