Isoforms of Cathepsin B1 in Neurotropic Schistosomula of
Trichobilharzia regenti Differ in Substrate Preferences and a
Highly Expressed Catalytically Inactive Paralog Binds Cystatin

J 2020

Isoforms of Cathepsin B1 in Neurotropic Schistosomula of Trichobilharzia regenti Differ in Substrate Preferences and a Highly Expressed Catalytically Inactive Paralog Binds Cystatin

DVOŘÁKOVÁ, Hana, Roman LEONTOVYČ, Tomáš MACHÁČEK, Anthony J. O´DONOGHUE, Ondrej ŠEDO et. al.

Základní údaje

Originální název

Isoforms of Cathepsin B1 in Neurotropic Schistosomula of Trichobilharzia regenti Differ in Substrate Preferences and a Highly Expressed Catalytically Inactive Paralog Binds Cystatin

Autoři

DVOŘÁKOVÁ, Hana (203 Česká republika), Roman LEONTOVYČ (203 Česká republika), Tomáš MACHÁČEK (203 Česká republika), Anthony J. O´DONOGHUE (840 Spojené státy), Ondrej ŠEDO (203 Česká republika, domácí), Zbyněk ZDRÁHAL (203 Česká republika, garant, domácí), Charles S. CRAIK (840 Spojené státy), Conor R. CAFFREY (840 Spojené státy), Petr HORÁK (203 Česká republika) a Libor MIKEŠ (203 Česká republika)

Vydání

FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, 2020, 2235-2988

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10606 Microbiology

Stát vydavatele

Švýcarsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

fulltext

Impakt faktor

Impact factor: 5.293

Kód RIV

RIV/00216224:14740/20:00115570

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.3389/fcimb.2020.00066

UT WoS

000524706100001

Klíčová slova anglicky

peptidase; cathepsin B; processing; substrate specificity; occluding loop; cystatin; helminth; schistosome

Štítky

CF PROT, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 2. 11. 2024 20:32, Ing. Martina Blahová

Anotace

V originále

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.

Návaznosti

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CEITEC 2020

90043, velká výzkumná infrastruktura

Název: CIISB

90127, velká výzkumná infrastruktura

Název: CIISB II

Citovat

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. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY. 2020, roč. 10, FEB, s. "66", 17 s. ISSN 2235-2988. Dostupné z: https://dx.doi.org/10.3389/fcimb.2020.00066.

@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}. 2020, roč.~10, FEB, s.~''66'', 17 s. ISSN~2235-2988. Dostupné z: https://dx.doi.org/10.3389/fcimb.2020.00066.

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