High selectivity of the hyperthermophilic subtilase propeptide domain toward inhibition of its cognate protease

Bahun, Miha (avtor), Poklar Ulrih, Nataša (avtor)

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.

• Jezik: Angleški jezik
• Ključne besede: hyperthermophile, subtilase, propeptide, enzyme activation, enzyme inhibition, protein stability
• Vrsta gradiva: Znanstveno delo
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: BF - Biotehniška fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2023
• Št. strani: 24 str.
• Številčenje: Vol. 11, iss. 5, art. 01487-23
• PID: 20.500.12556/RUL-155313
• UDK: 577.15
• ISSN pri članku: 2165-0497
• DOI: 10.1128/spectrum.01487-23
• COBISS.SI-ID: 163687939
• Datum objave v RUL: 26.03.2024

Gradivo je del revije

• Naslov: Microbiology spectrum
• Skrajšan naslov: Microbiol. spectr.
• Založnik: ASM Press
• ISSN: 2165-0497
• COBISS.SI-ID: 4526968

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: hipertermofili, encimi, proteaze

Projekti

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: P4-0121
• Naslov: Biokemijska in biofizikalno-kemijska karakterizacija naravnih snovi