20.500.12556/RUL-155313 High selectivity of the hyperthermophilic subtilase propeptide domain toward inhibition of its cognate protease Microbial extracellular subtilases are highly active proteolytic enzymes commonly used in commercial applications. These subtilases are synthesized in their inactive proform, which matures into the active protease under the control of the propeptide domain. In mesophilic bacterial prosubtilases, the propeptide functions as both an obligatory chaperone and an inhibitor of the subtilase catalytic domain. In contrast, the propeptides of hyperthermophilic archaeal prosubtilases act mainly as tight inhibitors and are not essential for subtilase folding. It is unclear whether this stronger inhibitory activity of hyperthermophilic propeptides results in their higher selectivity toward their cognate subtilases, in contrast to promiscuous mesophilic propeptides. Here, we showed that the propeptide of pernisine, a hyperthermostable archaeal subtilase, strongly interacts with and inhibits pernisine, but not the homol­ogous subtilisin Carlsberg and proteinase K. Instead, the pernisine propeptide was readily degraded by subtilisin Carlsberg and proteinase K. In addition, the catalytic domain of unprocessed propernisine was also susceptible to degradation but became proteolytically stable after autoprocessing of propernisine into the inactive, noncovalent complex propeptide:pernisine. This allowed efficient transactivation of the autopro­cessed complex propeptide:pernisine through degradation of pernisine propeptide by subtilisin Carlsberg and proteinase K at mesophilic temperature. Moreover, we demon­strated that active pernisine molecules are inhibited by the propeptide that is released after pernisine-catalyzed degradation of the unprocessed propernisine catalytic domain. This highlights the high inhibitory potency of the hyperthermophilic propeptide toward its cognate subtilase and its importance in regulating subtilase maturation, to prevent the degradation of the unprocessed subtilase precursors by the prematurely activated molecules. hyperthermophile subtilase propeptide enzyme activation enzyme inhibition protein stability hipertermofili encimi proteaze true false true Angleški jezik Slovenski jezik Znanstveno delo 2024-03-26 08:01:39 2024-03-26 08:01:45 2024-03-27 03:45:32 0000-00-00 00:00:00 2023 0 0 24 str. iss. 5, art. 01487-23 Vol. 11 Sep./Oct. 2023 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.15 2165-0497 10.1128/spectrum.01487-23 163687939 id_163687939_bahun-et-al-2023-high-selectivity-of-the-hyperthermophilic-subtilase-propeptide-domain-toward-inhibition-of-its-cognate.pdf id_163687939_bahun-et-al-2023-high-selectivity-of-the-hyperthermophilic-subtilase-propeptide-domain-toward-inhibition-of-its-cognate.pdf 1 8C644DAAD0688520422365ED19425043 fbbb6d587c29284b9a10dbcea5fba234dd98eb05189f2dd54b217a2f4519aebb 20232733-eb40-11ee-ab4b-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=182143 https://journals.asm.org/doi/10.1128/spectrum.01487-23 1 b4d6f08b-eb3e-11ee-ab4b-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=182142 Biotehniška fakulteta 0 0 0