A group of cathepsins L as predominant proteolytic enzymes of
Eudiplozoon nipponicum

a 2017

A group of cathepsins L as predominant proteolytic enzymes of Eudiplozoon nipponicum

JEDLIČKOVÁ, Lucie; Hana DVOŘÁKOVÁ; Jan DVOŘÁK; John P. DALTON; Martin KAŠNÝ et al.

Základní údaje

Originální název

A group of cathepsins L as predominant proteolytic enzymes of Eudiplozoon nipponicum

Autoři

JEDLIČKOVÁ, Lucie; Hana DVOŘÁKOVÁ; Jan DVOŘÁK; John P. DALTON; Martin KAŠNÝ; Lenka ULRYCHOVÁ; Jiří VOREL a Libor MIKEŠ

Vydání

8th International Symposium on Monogenea, 2017

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10600 1.6 Biological sciences

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

ECIP - European Centre of IchthyoParasitology

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/17:00094960

Organizační jednotka

Přírodovědecká fakulta

ISBN

978-80-210-8666-1

Klíčová slova anglicky

Sequencing; transcriptomics; helminths; Eudiplozoon nipponicum; Monogenea

Příznaky

Mezinárodní význam

Změněno: 20. 9. 2017 14:23, Mgr. Jiří Vorel, Ph.D.

Anotace

V originále

Cathepsin L-like peptidases of the hematophagous monogenean Eudiplozoon nipponicum (family Diplozoidae) seem to play a key role in the digestion of host blood proteins. We have identified at least 13 different transcripts coding for cathepsins L in the transcriptome of adult worms. Two of them, the most transcribed ones, named EnCL1 [GenBank: KP793605] and EnCL3 [GenBank: KP793606], were heterologously expressed in E. coli bacterial (rEnCL1b) and P. pastoris yeast systems (rEnCL1y and rEnCL3y). While rEnCL3y was obtained as a relatively stable zymogen (proenzyme) and could be autoactivated at low pH, rEnCL1y undertook self-processing in yeast medium and was very unstable. Both recombinant enzymes exhibited substrate preferences characteristic for cathepsin L-like peptidases, and were able to hydrolyze hemoglobin, albumin, type I collagen, IgG, and fibrinogen. Specific RNA probes and monospecific antibodies localized the transcripts/enzymes inside the vesicles of haematin cells of the digestive tract. Moreover, both EnCL1 and EnCL3 were also detected in the gut lumen of adult worms. These results strongly support the idea that both CL endopeptidases studied participate in processing of blood. Additionally, bioinformatic sequence analyses were performed in order to compare the other cathepsin L-like sequences found in the transcriptome of adult E. nipponicum. One of the sequences (named EnCL2) has a conspicuously divergent composition of the S2 subsite, and could be expected to efficiently cleave the substrates with proline in P2 position, including the repeated Gly-Pro-Xaa motifs occurring within the amino acid sequence of collagen. Thanks to this, EnCL2 might participate in disruption of host tissues, e.g. of gill lamellae.

Návaznosti

GBP505/12/G112, projekt VaV

Název: ECIP - Evropské centrum ichtyoparazitologie

Investor: Grantová agentura ČR, ECIP - Evropské centrum ichtyoparazitologie

Citovat

JEDLIČKOVÁ, Lucie; Hana DVOŘÁKOVÁ; Jan DVOŘÁK; John P. DALTON; Martin KAŠNÝ; Lenka ULRYCHOVÁ; Jiří VOREL a Libor MIKEŠ. A group of cathepsins L as predominant proteolytic enzymes of Eudiplozoon nipponicum. In 8th International Symposium on Monogenea. 2017. ISBN 978-80-210-8666-1.

@proceedings{1390125,
   author = {Jedličková, Lucie and Dvořáková, Hana and Dvořák, Jan and Dalton, John P. and Kašný, Martin and Ulrychová, Lenka and Vorel, Jiří and Mikeš, Libor},
   booktitle = {8th International Symposium on Monogenea},
   keywords = {Sequencing; transcriptomics; helminths; Eudiplozoon nipponicum; Monogenea},
   language = {eng},
   isbn = {978-80-210-8666-1},
   title = {A group of cathepsins L as predominant proteolytic enzymes of Eudiplozoon nipponicum},
   url = {http://ecip.cz/news/4},
   year = {2017}
}

TY  - CONF
ID  - 1390125
AU  - Jedličková, Lucie - Dvořáková, Hana - Dvořák, Jan - Dalton, John P. - Kašný, Martin - Ulrychová, Lenka - Vorel, Jiří - Mikeš, Libor
PY  - 2017
TI  - A group of cathepsins L as predominant proteolytic enzymes of Eudiplozoon nipponicum
SN  - 9788021086661
KW  - Sequencing
KW  - transcriptomics
KW  - helminths
KW  - Eudiplozoon nipponicum
KW  - Monogenea
UR  - http://ecip.cz/news/4
L2  - http://ecip.cz/news/4
N2  - Cathepsin L-like peptidases of the hematophagous monogenean Eudiplozoon nipponicum (family Diplozoidae) seem to play a key role in the digestion of host blood proteins. We have identified at least 13 different transcripts coding for cathepsins L in the transcriptome of adult worms. Two of them, the most transcribed ones, named EnCL1 [GenBank: KP793605] and EnCL3 [GenBank: KP793606], were heterologously expressed in E. coli bacterial (rEnCL1b) and P. pastoris yeast systems (rEnCL1y and rEnCL3y). While rEnCL3y was obtained as a relatively stable zymogen (proenzyme) and could be autoactivated at low pH, rEnCL1y undertook self-processing in yeast medium and was very unstable. Both recombinant enzymes exhibited substrate preferences characteristic for cathepsin L-like peptidases, and were able to hydrolyze hemoglobin, albumin, type I collagen, IgG, and fibrinogen. Specific RNA probes and monospecific antibodies localized the transcripts/enzymes inside the vesicles of haematin cells of the digestive tract. Moreover, both EnCL1 and EnCL3 were also detected in the gut lumen of adult worms. These results strongly support the idea that both CL endopeptidases studied participate in processing of blood. Additionally, bioinformatic sequence analyses were performed in order to compare the other cathepsin L-like sequences found in the transcriptome of adult E. nipponicum. One of the sequences (named EnCL2) has a conspicuously divergent composition of the S2 subsite, and could be expected to efficiently cleave the substrates with proline in P2 position, including the repeated Gly-Pro-Xaa motifs occurring within the amino acid sequence of collagen. Thanks to this, EnCL2 might participate in disruption of host tissues, e.g. of gill lamellae.
ER  -

JEDLIČKOVÁ, Lucie; Hana DVOŘÁKOVÁ; Jan DVOŘÁK; John P. DALTON; Martin KAŠNÝ; Lenka ULRYCHOVÁ; Jiří VOREL a Libor MIKEŠ. A group of cathepsins L as predominant proteolytic enzymes of Eudiplozoon nipponicum. In \textit{8th International Symposium on Monogenea}. 2017. ISBN~978-80-210-8666-1.

Přehled o publikaci