Cysteine peptidases of Eudiplozoon nipponicum: a broad repertoire of structurally assorted cathepsins L in contrast to the scarcity of cathepsins B in an invasive species of haematophagous monogenean of common carp

Jedličková, Lucie; Dvořáková, Hana; Dvořák, Jan; Kašný, Martin; Ulrychová, Lenka; Vorel, Jiří; Žárský, Vojtěch; Mikeš, Libor

doi:10.1186/s13071-018-2666-2

Cited by 26 publications

(39 citation statements)

References 79 publications

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“…This can be done repeatedly and for different proteins without the need for additional experiments. The results of this study show that this method confirmed the localization of previously characterized cathepsins and kunitz of E. nipponicum [29,42] (Table 3). On the other hand, there is still room for improvement because two of the eleven examined cathepsins were not identified and another two were identified in tissues where they were not expected.…”

Section: Discussionsupporting

confidence: 85%

“…Moreover, functional proteins from the intestine are of great importance due to their involvement in host-parasite interplay and a better understanding of these processes could help contribute to a control of infection not only by E. nipponicum but possibly other monogeneans as well. However, only a handful of studies so far focused on particular functional proteins [29,30,42–44] and the more complex proteomic data have been missing until now.…”

Section: Discussionmentioning

confidence: 99%

“…In terms of tissue specificity of peptidases and inhibitors (Fig 4 and 6), the richest environment is the intestine, which is the most metabolically active: both digestion and nutrient uptake take place in the gut lumen and hematin cells of the gastrodermis [29,52]. The tegument ended up the second, which could be due to the fact that this tissue protects the rest of the body from the environment.…”

Section: Discussionmentioning

confidence: 99%

“…In all these statistics, the intestine was once again dominant with respect to variety. This is once again related to its function, namely feeding, since most peptidases and inhibitors play a role in digestion, inhibition of blood coagulation, and prevention of inflammation [29,42,52].…”

Section: Discussionmentioning

confidence: 99%

“…In recent years, this situation started to improve because E. nipponicum has been studied in a broader context. Several functional protein molecules of E. nipponicum were described [29,30,41–44] and the genome, transcriptome, and secretome of this organism are soon to be published. With this study, we significantly enrich available information on monogenean functional molecular biology by describing protein distribution in selected E. nipponicum tissues.…”

Section: Introductionmentioning

confidence: 99%

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Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes ofEudiplozoon nipponicum(Monogenea, Polyopisthocotylea)

Roudnický

Potěšil²,

Zdráhal³

et al. 2020

Preprint

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23Eudiplozoon nipponicum (Goto, 1891) is a hematophagous monogenean ectoparasite 24 which inhabits the gills of the common carp (Cyprinus carpio). Heavy infestation can lead to 25 anemia and in conjunction with secondary bacterial infections cause poor health and eventual 26 death of the host. 27 This study is based on an innovative approach to protein localization which has never 28 been used in parasitology before. Using laser capture microdissection, we dissected particular 29 areas of the parasite body without contaminating the samples by surrounding tissue, and in 30 combination with analysis by mass spectrometry obtained tissue-specific proteomes of 31 tegument, intestine, and parenchyma of our model organism, E. nipponicum. We successfully 32 verified the presence of certain functional proteins (e.g. cathepsin L) in tissues where their 33 presence was expected (intestine) and confirmed that there were no traces of these proteins in 34 other tissues (tegument and parenchyma). Additionally, we identified a total of 2,059 proteins, 35 including 72 peptidases and 33 peptidase inhibitors. As expected, the greatest variety was found 36 in the intestine and the lowest variety in the parenchyma. 37Our results are significant on two levels. Firstly, we demonstrated how one can localize 38 all proteins in one analysis and without using laboratory animals (antibodies for 39 immunolocalization of single proteins). Secondly, this study offers the first complex proteomic 40 data on not only the E. nipponicum but within the whole class of Monogenea, which was from 41 this point of view until recently neglected. 42 3 43 Introduction 44 Laser-capture microdissection (LCM) was developed as a powerful and reliable tool to 45 overcome the heterogeneity of the specimen [1]. It enables isolation of specific tissues, 46 individual cells, or even individual organelles from complex samples based on cell morphology 47 [2,3]. The method was first described in the second half of the 20 th century and since then further 48 developed and modified [2,4-7]. Nowadays, LCM combines laser excision with high-resolution 49 microscopic control, which makes it possible to precisely capture for instance individual cells 50 or even only nucleus while keeping track of the location and morphology of the source tissue 51 [2,3,8]. 52 LCM technology has been used in a wide variety of applications with focus on genomic, 53 transcriptomic, and even proteomic analyses, such as dissection of polar bodies from oocytes 54 for pre-fertilization genetic diagnosis [9], transcriptome-wide analysis of blood vessels from 55 human skin and wound-edge tissue [10], proteomic profiling of dentoalveolar tissues [11], and 56 many other areas [12-14]. But only one study so far used a combination of LCM and mass 57 spectrometry (MS) to localize unique proteins, potential biomarkers, when dealing with the 58 heterogeneity of breast tumor [15]. 59 LCM has also been used in parasitology, especially in sample preparation. Recent 60 studies employed this method in...

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes ofEudiplozoon nipponicum(Monogenea, Polyopisthocotylea)

Roudnický

Potěšil²,

Zdráhal³

et al. 2020

Preprint

Self Cite

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The Nϵ‐Rule for Serine, but Not Cysteine Catalytic Triads

Czapinska

Bochtler

2022

Angewandte Chemie

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Catalytic triads, composed of a serine or cysteine nucleophile, a histidine, and a third triad residue (typically Asp/Glu/Asn), are common in enzyme active sites and catalyze a wide variety of chemical reactions. Two types of triads can be distinguished: We refer to them as Nδ-or Nɛ-configured, depending on whether the histidine imidazole Nδ or Nɛ atom is close to the nucleophile Oγ/Sγ. In this study, we have analyzed triad configuration. In structural triads, the more stable Nδ-configuration predominates. For catalytic triads, the configuration depends on the nucleophile. When it is a cysteine residue, both configuration types occur, depending on the family. However, when the nucleophile is a serine residue, the less stable Nɛ-configuration is almost exclusively found. We posit that the energetically less favored conformation is selected for in serine triads to facilitate the otherwise difficult proton transfer from the nucleophile to the histidine residue.

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