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@article{1649841,
author = {Dvořáková, Hana and Leontovyč, Roman and Macháček, Tomáš and O´Donoghue, Anthony J. and Šedo, Ondrej and Zdráhal, Zbyněk and Craik, Charles S. and Caffrey, Conor R. and Horák, Petr and Mikeš, Libor},
article_number = {FEB},
doi = {http://dx.doi.org/10.3389/fcimb.2020.00066},
keywords = {peptidase; cathepsin B; processing; substrate specificity; occluding loop; cystatin; helminth; schistosome},
language = {eng},
issn = {2235-2988},
journal = {FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY},
title = {Isoforms of Cathepsin B1 in Neurotropic Schistosomula of Trichobilharzia regenti Differ in Substrate Preferences and a Highly Expressed Catalytically Inactive Paralog Binds Cystatin},
url = {https://www.frontiersin.org/articles/10.3389/fcimb.2020.00066/full},
volume = {10},
year = {2020}
}
TY - JOUR
ID - 1649841
AU - Dvořáková, Hana - Leontovyč, Roman - Macháček, Tomáš - O´Donoghue, Anthony J. - Šedo, Ondrej - Zdráhal, Zbyněk - Craik, Charles S. - Caffrey, Conor R. - Horák, Petr - Mikeš, Libor
PY - 2020
TI - Isoforms of Cathepsin B1 in Neurotropic Schistosomula of Trichobilharzia regenti Differ in Substrate Preferences and a Highly Expressed Catalytically Inactive Paralog Binds Cystatin
JF - FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
VL - 10
IS - FEB
SP - "66"
EP - "66"
SN - 22352988
KW - peptidase
KW - cathepsin B
KW - processing
KW - substrate specificity
KW - occluding loop
KW - cystatin
KW - helminth
KW - schistosome
UR - https://www.frontiersin.org/articles/10.3389/fcimb.2020.00066/full
L2 - https://www.frontiersin.org/articles/10.3389/fcimb.2020.00066/full
N2 - Schistosomula (the post-infective stages) of the neurotropic schistosome Trichobilharzia regenti possess multiple isoforms of cathepsin B1 peptidase (TrCB1.1-TrCB1.6) with involvement in nutrient digestion. The comparison of substrate preferences of TrCB1.1 and TrCB1.4 showed that TrCB1.4 had a very narrow substrate specificity and after processing it was less effective toward protein substrates when compared to TrCB1.1. Self-processing of both isoforms could be facilitated by sulfated polysaccharides due to a specific binding motif in the pro-sequence. Trans-activation by heterologous enzymes was also successfully employed. Expression profiling revealed a high level of transcription of genes encoding the enzymatically inactive paralogs TrCB1.5 and TrCB1.6. The transcription level of TrCB1.6 was comparable with that of TrCB1.1 and TrCB1.2, the most abundant active isoforms. Recombinant TrCB1.6wt, a wild type paralog with a Cys(29)-to-Gly substitution in the active site that renders the enzyme inactive, was processed by the active TrCB1 forms and by an asparaginyl endopeptidase. Although TrCB1.6wt lacked hydrolytic activity, endopeptidase, but not dipeptidase, activity could be restored by mutating Gly(29) to Cys(29). The lack of exopeptidase activity may be due to other mutations, such as His(110)-to-Asn in the occluding loop and Asp(224)-to-Gly in the main body of the mature TrCB1.6, which do not occur in the active isoforms TrCB1.1 and TrCB1.4 with exopeptidase activity. The catalytically active enzymes and the inactive TrCB1.6 paralog formed complexes with chicken cystatin, thus supporting experimentally the hypothesis that inactive paralogs could potentially regulate the activity of the active forms or protect them from being inhibited by host inhibitors. The effect on cell viability and nitric oxide production by selected immune cells observed for TrCB1.1 was not confirmed for TrCB1.6. We show here that the active isoforms of TrCB1 have different affinities for peptide substrates thereby facilitating diversity in protein-derived nutrition for the parasite. The inactive paralogs are unexpectedly highly expressed and one of them retains the ability to bind cystatins, likely due to specific mutations in the occluding loop and the enzyme body. This suggests a role in sequestration of inhibitors and protection of active cysteine peptidases.
ER -
DVOŘÁKOVÁ, Hana, Roman LEONTOVYČ, Tomáš MACHÁČEK, Anthony J. O´DONOGHUE, Ondrej ŠEDO, Zbyněk ZDRÁHAL, Charles S. CRAIK, Conor R. CAFFREY, Petr HORÁK a Libor MIKEŠ. Isoforms of Cathepsin B1 in Neurotropic Schistosomula of Trichobilharzia regenti Differ in Substrate Preferences and a Highly Expressed Catalytically Inactive Paralog Binds Cystatin. \textit{FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY}. roč.~10, FEB, s.~''66'', 17 s. ISSN~2235-2988. doi:10.3389/fcimb.2020.00066. 2020.
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