Characterization of Gut-associated Cathepsin D Hemoglobinase from Tick Ixodes ricinus (IrCD1)

Sojka, Daniel; Franta, Zdeněk; Frantová, Helena; Bartošová, Pavla; Horn, Martin; Váchová, Jana; O’Donoghue, A. J.; Eroy-Reveles, A. A.; Craik, C. S.; Knudsen, G. M.; Caffrey, C. R.; McKerrow, J. H.; Mareš, Michael; Kopáček, Petr

doi:https://dx.doi.org/10.1074/jbc.M112.347922

Number of the records: 1

Characterization of Gut-associated Cathepsin D Hemoglobinase from Tick Ixodes ricinus (IrCD1)

1.

SYSNO ASEP	0377766
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Characterization of Gut-associated Cathepsin D Hemoglobinase from Tick Ixodes ricinus (IrCD1)
Author(s)	Sojka, Daniel (BC-A)_{RID, ORCID} Franta, Zdeněk (BC-A) Frantová, Helena (BC-A)_ORCID Bartošová, Pavla (BC-A)_{RID, ORCID} Horn, Martin (UOCHB-X)_{RID, ORCID} Váchová, Jana (UOCHB-X) O’Donoghue, A. J. (US) Eroy-Reveles, A. A. (US) Craik, C. S. (US) Knudsen, G. M. (US) Caffrey, C. R. (US) McKerrow, J. H. (US) Mareš, Michael (UOCHB-X)_{RID, ORCID} Kopáček, Petr (BC-A)_{RID, ORCID}
Source Title	Journal of Biological Chemistry. - : Elsevier - ISSN 0021-9258 Roč. 287, č. 25 (2012), s. 21152-21163
Number of pages	12 s.
Language	eng - English
Country	US - United States
Keywords	RHIPICEPHALUS BOOPHILUS MICROPLUS ; HARD TICK ; ORNITHODOROS-MOUBATA ; ASPARTIC PROTEINASE ; RECOGNITION DOMAIN ; MOLECULAR-CLONING ; ACTIVE-SITE ; EXPRESSION ; INHIBITOR ; PROTEASE
Subject RIV	EB - Genetics ; Molecular Biology
R&D Projects	IAA600960910 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR)
	LC06009 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	KJB600960911 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR)
	GPP502/11/P682 GA ČR - Czech Science Foundation (CSF)
	IAA400550705 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR)
	GAP207/10/2183 GA ČR - Czech Science Foundation (CSF)
Institutional support	UOCHB-X - RVO:61388963
CEZ	AV0Z60220518 - PAU-O, BC-A (2005-2011)
UT WOS	000306416800036
DOI	10.1074/jbc.M112.347922
Annotation	To identify the gut-associated tick aspartic hemoglobinase, this work focuses on the functional diversity of multiple Ixodes ricinus cathepsin D forms (IrCDs). Out of three encoding genes representing Ixodes scapularis genome paralogs, IrCD1 is the most distinct enzyme with a shortened propeptide region and a unique pattern of predicted post-translational modifications. IrCD1 gene transcription is induced by tick feeding and is restricted to the gut tissue. The hemoglobinolytic role of IrCD1 was further supported by immunolocalization of IrCD1 in the vesicles of tick gut cells. Properties of recombinantly expressed rIrCD1 are consistent with the endo-lysosomal environment because the zymogen is autoactivated and remains optimally active in acidic conditions. Hemoglobin cleavage pattern of rIrCD1 is identical to that produced by the native enzyme. The preference for hydrophobic residues at the P1 and P1' position was confirmed by screening a novel synthetic tetradecapeptidyl substrate library. Outside the S1-S1' regions, rIrCD1 tolerates most amino acids but displays a preference for tyrosine at P3 and alanine at P2'. Further analysis of the cleavage site location within the peptide substrate indicated that IrCD1 is a true endopeptidase. The role in hemoglobinolysis was verified with RNAi knockdown of IrCD1 that decreased gut extract cathepsin D activity by >90%. IrCD1 was newly characterized as a unique hemoglobinolytic cathepsin D contributing to the complex intestinal proteolytic network of mainly cysteine peptidases in ticks.
Workplace	Biology Centre (since 2006)
Contact	Dana Hypšová, eje@eje.cz, Tel.: 387 775 214
Year of Publishing	2013

Number of the records: 1