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Projects / Programmes source: ARIS

MECHANISM OF LIPID MEMBRANE DAMAGE INDUCED BY Nep1-LIKE PROTEINS

Research activity

Code Science Field Subfield
1.05.00  Natural sciences and mathematics  Biochemistry and molecular biology   
1.03.02  Natural sciences and mathematics  Biology  Botany 

Code Science Field
B230  Biomedical sciences  Microbiology, bacteriology, virology, mycology 

Code Science Field
1.06  Natural Sciences  Biological sciences 
Keywords
Nep1-like proteins; microbial pathogenesis; lipid membranes; lipid membrane damage; nanopores
Evaluation (rules)
source: COBISS
Researchers (17)
no. Code Name and surname Research area Role Period No. of publicationsNo. of publications
1.  15686  PhD Gregor Anderluh  Biochemistry and molecular biology  Head  2019 - 2022  970 
2.  24290  PhD Matej Butala  Biochemistry and molecular biology  Researcher  2022  236 
3.  53283  Maja Hostnik  Biochemistry and molecular biology  Researcher  2022  19 
4.  53439  PhD Magdalena Kulma  Biochemistry and molecular biology  Researcher  2021  32 
5.  33923  PhD Polona Mrak  Natural sciences and mathematics  Researcher  2019 - 2022  102 
6.  39090  PhD Anastasija Panevska  Biochemistry and molecular biology  Researcher  2019 - 2021  57 
7.  30762  PhD Katja Pirc  Biochemistry and molecular biology  Researcher  2021 - 2022  46 
8.  12048  PhD Marjetka Podobnik  Biochemistry and molecular biology  Researcher  2019 - 2022  321 
9.  15328  PhD Kristina Sepčić  Biochemistry and molecular biology  Researcher  2019 - 2021  735 
10.  33137  PhD Matej Skočaj  Biochemistry and molecular biology  Researcher  2019 - 2022  109 
11.  50612  Tina Snoj  Biochemistry and molecular biology  Junior researcher  2019 - 2021  19 
12.  53732  Marija Srnko  Biochemistry and molecular biology  Researcher  2019 - 2022  22 
13.  38479  PhD Aleksandra Šakanović  Neurobiology  Researcher  2019 - 2020  25 
14.  38473  Tomaž Švigelj    Technical associate  2019 - 2022  11 
15.  21684  Tea Tomšič    Technical associate  2019 - 2020 
16.  06905  PhD Tom Turk  Biochemistry and molecular biology  Researcher  2021  620 
17.  16381  PhD Nada Žnidaršič  Biology  Researcher  2019 - 2022  243 
Organisations (2)
no. Code Research organisation City Registration number No. of publicationsNo. of publications
1.  0104  National Institute of Chemistry  Ljubljana  5051592000  21,092 
2.  0481  University of Ljubljana, Biotechnical Faculty  Ljubljana  1626914  66,958 
Abstract
Fight against pathogens and antimicrobial resistance is a big and pressing problem of modern society. Novel strategies and targets for development of antimicrobial substances are thus crucially needed in areas as diverse as medicine, production of safe food, etc. The members of the family of necrosis- and ethylene-inducing peptide 1 (NEP1)-like proteins, i.e. NLPs, elicit diverse defence reactions and cell death in eudicot plants but not in monocots. NLPs are widely distributed among taxonomically nonrelated microorganisms like fungi, bacteria and oomycetes. These microorganisms are widespread, they may infect range of different crops, such as potato, tomato, soya and tobacco, and cause enormous economic loss worldwide. It was shown that NLPs function as cytolytic toxins that induce plasma membrane leakage, thus causing cytotoxicity. Based on their crystal structures, NLPs are considered to be distantly related to other pore-forming toxins of animal origin, such as actinoporins from sea anemones. The mechanism by which NLP induce necrosis is poorly understood. Recently, we have identified glycosylinositol phosphorylceramides (GIPC), a major class of plant sphingolipids, as a target molecule for NLP binding to plant plasma membranes (Lenarčič et al., Science, 2017). GIPCs consist of a polar headgroup bearing variable carbohydrate moieties and inositol phosphorylceramide core. Type and number of terminal hexose groups varies significantly between plant species and plant tissues. Binding of the GIPC terminal hexose moiety induces several conformational changes within the NLP toxin that may precede membrane attachment and host cell lysis. Nothing is known about other steps of membrane damage subsequent to membrane binding and this project aims to clarify steps of membrane damage induced by NLP proteins. We will test the central hypothesis of the project that NLPs damage lipid membranes by a multistep process that lead to pore formation. The main objectives of this project are therefore to (i) determine the molecular mechanism of membrane damage by NLPs, (ii) resolve underlying selectivity of NLPs against eudicot plants at the molecular level and (iii) determine structural features of oligomeric assemblies of NLPs formed at the surface of lipid membranes composed of GIPCs. To achieve objectives of this project proposal we will use state-of-the-art biochemical, biophysical, molecular and structural biology approaches, including X-ray crystallography and cryo-electron microscopy, and molecular modelling.   The results of this project will provide molecular details of NLPs interactions with target cell surface, molecular mechanism of membrane damage and properties of pores formed by NLPs. This will clarify steps in pathogenesis of some of the most pressing pathogens and open avenues for development of strategies for fighting against microbial pathogens.
Significance for science
Antimicrobial resistance is a big and pressing problem in medicine and other areas such as production of safe food. This project will importantly contribute to the fight against pathogenic microorganisms by clarifying basic steps in pathogenesis of some of the most important pathogenic microorganisms. The description of the NLP pore at high resolution will allow designing novel approaches for fighting pathogenic microorganisms. High-resolution structures are prerequisite for rational design of inhibitory substances that will specifically act upon NLP proteins and cause little damage to other organisms or environment. This will be important contribution to the fight against pathogenic microorganisms. Since NLP proteins are widely distributed and highly similar in structure they are perfect example for such approaches and development of antimicrobial substances. This project will allow clarification of membrane-damage mechanism induced by NLP proteins. These proteins are present in many different pathogenic microorganisms and have crucial role in pathogenesis. They act upon many agriculturally important crops such as potato or tomato. The results of this study will importantly contribute to our understanding of NLP proteins interactions with the surface of plant cells, the mechanism of membrane damage induced by NLP proteins and properties of pores formed by NLP proteins. We expect that we will be able to describe NLP pores at high resolution. Since the structure of NLP proteins is different from other cytolytic protein families it is reasonable to expect that we will reveal a novel mechanism of membrane damage. New nanopores will be employed in sensing applications and will thus also allow novel approaches in biosensing.
Significance for the country
Antimicrobial resistance is a big and pressing problem in medicine and other areas such as production of safe food. This project will importantly contribute to the fight against pathogenic microorganisms by clarifying basic steps in pathogenesis of some of the most important pathogenic microorganisms. The description of the NLP pore at high resolution will allow designing novel approaches for fighting pathogenic microorganisms. High-resolution structures are prerequisite for rational design of inhibitory substances that will specifically act upon NLP proteins and cause little damage to other organisms or environment. This will be important contribution to the fight against pathogenic microorganisms. Since NLP proteins are widely distributed and highly similar in structure they are perfect example for such approaches and development of antimicrobial substances. This project will allow clarification of membrane-damage mechanism induced by NLP proteins. These proteins are present in many different pathogenic microorganisms and have crucial role in pathogenesis. They act upon many agriculturally important crops such as potato or tomato. The results of this study will importantly contribute to our understanding of NLP proteins interactions with the surface of plant cells, the mechanism of membrane damage induced by NLP proteins and properties of pores formed by NLP proteins. We expect that we will be able to describe NLP pores at high resolution. Since the structure of NLP proteins is different from other cytolytic protein families it is reasonable to expect that we will reveal a novel mechanism of membrane damage. New nanopores will be employed in sensing applications and will thus also allow novel approaches in biosensing.
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