A novel Kunitz protein with proposed dual function from
Eudiplozoon nipponicum (Monogenea) impairs haemostasis and
action of complement in vitro

J 2019

A novel Kunitz protein with proposed dual function from Eudiplozoon nipponicum (Monogenea) impairs haemostasis and action of complement in vitro

JEDLIČKOVÁ, Lucie, Jan DVOŘÁK, Ingrid HRACHOVINOVÁ, Lenka ULRYCHOVÁ, Martin KAŠNÝ et. al.

Basic information

Original name

A novel Kunitz protein with proposed dual function from Eudiplozoon nipponicum (Monogenea) impairs haemostasis and action of complement in vitro

Authors

JEDLIČKOVÁ, Lucie (203 Czech Republic), Jan DVOŘÁK (203 Czech Republic), Ingrid HRACHOVINOVÁ (203 Czech Republic), Lenka ULRYCHOVÁ (203 Czech Republic), Martin KAŠNÝ (203 Czech Republic, guarantor, belonging to the institution) and Libor MIKEŠ (203 Czech Republic)

Edition

International Journal for Parasitology, Oxford, Elsevier Science Ltd. 2019, 0020-7519

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10613 Zoology

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.530

RIV identification code

RIV/00216224:14310/19:00108143

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1016/j.ijpara.2018.11.010

UT WoS

000466453400002

Keywords in English

Anticoagulant; Kunitz; Peptidase inhibitor; Factor X; Plasmin; Haematophagy; Helminth; Diplozoidae

Abstract

V originále

Serine peptidases are involved in many physiological processes including digestion, haemostasis and complement cascade. Parasites regulate activities of host serine peptidases to their own benefit, employing various inhibitors, many of which belong to the Kunitz-type protein family. In this study, we confirmed the presence of potential anticoagulants in protein extracts of the haematophagous monogenean Eudiplozoon nipponicum which parasitizes the common carp. We then focused on a Kunitz protein (EnKT1) discovered in the E. nipponicum transcriptome, which structurally resembles textilinin-1, an antihemorrhagic snake venom factor from Pseudonaja textilis. The protein was recombinantly expressed, purified and biochemically characterised. The recombinant EnKT1 did inhibit in vitro activity of Factor Xa of the coagulation cascade, but exhibited a higher activity against plasmin and plasma kallikrein, which participate in fibrinolysis, production of kinins, and complement activation. Anti-coagulation properties of EnKT1 based on the inhibition of Factor Xa were confirmed by thromboe-lastography, but no effect on fibrinolysis was observed. Moreover, we discovered that EnKT1 significantly impairs the function of fish complement, possibly by inhibiting plasmin or Factor Xa which can act as a C3 and C5 convertase. We localised Enkt1 transcripts and protein within haematin digestive cells of the parasite by RNA in situ hybridisation and immunohistochemistry, respectively. Based on these results, we suggest that the secretory Kunitz protein of E. nipponicum has a dual function. In particular, it impairs both haemostasis and complement activation in vitro, and thus might facilitate digestion of a host's blood and protect a parasite's gastrodermis from damage by the complement. This study presents, to our knowledge, the first characterisation of a Kunitz protein from monogeneans and the first example of a parasite Kunitz inhibitor that impairs the function of the complement. (C) 2019 The Author(s). Published by Elsevier Ltd on behalf of Australian Society for Parasitology.

Links

GAP506/12/1258, research and development project

Name: Interakce hostitel-parazit u krevsajících diplozoidních monogeneí: Výzkum vysoce specializovaných adaptací k parazitismu

Investor: Czech Science Foundation

GBP505/12/G112, research and development project

Name: ECIP - Evropské centrum ichtyoparazitologie

Investor: Czech Science Foundation

LM2015043, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

JEDLIČKOVÁ, Lucie, Jan DVOŘÁK, Ingrid HRACHOVINOVÁ, Lenka ULRYCHOVÁ, Martin KAŠNÝ and Libor MIKEŠ. A novel Kunitz protein with proposed dual function from Eudiplozoon nipponicum (Monogenea) impairs haemostasis and action of complement in vitro. International Journal for Parasitology. Oxford: Elsevier Science Ltd., 2019, vol. 49, No 5, p. 337-346. ISSN 0020-7519. Available from: https://dx.doi.org/10.1016/j.ijpara.2018.11.010.

@article{1617117,
   author = {Jedličková, Lucie and Dvořák, Jan and Hrachovinová, Ingrid and Ulrychová, Lenka and Kašný, Martin and Mikeš, Libor},
   article_location = {Oxford},
   article_number = {5},
   doi = {http://dx.doi.org/10.1016/j.ijpara.2018.11.010},
   keywords = {Anticoagulant; Kunitz; Peptidase inhibitor; Factor X; Plasmin; Haematophagy; Helminth; Diplozoidae},
   language = {eng},
   issn = {0020-7519},
   journal = {International Journal for Parasitology},
   title = {A novel Kunitz protein with proposed dual function from Eudiplozoon nipponicum (Monogenea) impairs haemostasis and action of complement in vitro},
   url = {https://www.sciencedirect.com/science/article/pii/S0020751919300268?via%3Dihub},
   volume = {49},
   year = {2019}
}

TY  - JOUR
ID  - 1617117
AU  - Jedličková, Lucie - Dvořák, Jan - Hrachovinová, Ingrid - Ulrychová, Lenka - Kašný, Martin - Mikeš, Libor
PY  - 2019
TI  - A novel Kunitz protein with proposed dual function from Eudiplozoon nipponicum (Monogenea) impairs haemostasis and action of complement in vitro
JF  - International Journal for Parasitology
VL  - 49
IS  - 5
SP  - 337-346
EP  - 337-346
PB  - Elsevier Science Ltd.
SN  - 00207519
KW  - Anticoagulant
KW  - Kunitz
KW  - Peptidase inhibitor
KW  - Factor X
KW  - Plasmin
KW  - Haematophagy
KW  - Helminth
KW  - Diplozoidae
UR  - https://www.sciencedirect.com/science/article/pii/S0020751919300268?via%3Dihub
L2  - https://www.sciencedirect.com/science/article/pii/S0020751919300268?via%3Dihub
N2  - Serine peptidases are involved in many physiological processes including digestion, haemostasis and complement cascade. Parasites regulate activities of host serine peptidases to their own benefit, employing various inhibitors, many of which belong to the Kunitz-type protein family. In this study, we confirmed the presence of potential anticoagulants in protein extracts of the haematophagous monogenean Eudiplozoon nipponicum which parasitizes the common carp. We then focused on a Kunitz protein (EnKT1) discovered in the E. nipponicum transcriptome, which structurally resembles textilinin-1, an antihemorrhagic snake venom factor from Pseudonaja textilis. The protein was recombinantly expressed, purified and biochemically characterised. The recombinant EnKT1 did inhibit in vitro activity of Factor Xa of the coagulation cascade, but exhibited a higher activity against plasmin and plasma kallikrein, which participate in fibrinolysis, production of kinins, and complement activation. Anti-coagulation properties of EnKT1 based on the inhibition of Factor Xa were confirmed by thromboe-lastography, but no effect on fibrinolysis was observed. Moreover, we discovered that EnKT1 significantly impairs the function of fish complement, possibly by inhibiting plasmin or Factor Xa which can act as a C3 and C5 convertase. We localised Enkt1 transcripts and protein within haematin digestive cells of the parasite by RNA in situ hybridisation and immunohistochemistry, respectively. Based on these results, we suggest that the secretory Kunitz protein of E. nipponicum has a dual function. In particular, it impairs both haemostasis and complement activation in vitro, and thus might facilitate digestion of a host's blood and protect a parasite's gastrodermis from damage by the complement. This study presents, to our knowledge, the first characterisation of a Kunitz protein from monogeneans and the first example of a parasite Kunitz inhibitor that impairs the function of the complement. (C) 2019 The Author(s). Published by Elsevier Ltd on behalf of Australian Society for Parasitology.
ER  -

JEDLIČKOVÁ, Lucie, Jan DVOŘÁK, Ingrid HRACHOVINOVÁ, Lenka ULRYCHOVÁ, Martin KAŠNÝ and Libor MIKEŠ. A novel Kunitz protein with proposed dual function from Eudiplozoon nipponicum (Monogenea) impairs haemostasis and action of complement in vitro. \textit{International Journal for Parasitology}. Oxford: Elsevier Science Ltd., 2019, vol.~49, No~5, p.~337-346. ISSN~0020-7519. Available from: https://dx.doi.org/10.1016/j.ijpara.2018.11.010.

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