Calocera viscosa lectin – a fungi member of the jacalin-related
lectins family

k 2023

Calocera viscosa lectin – a fungi member of the jacalin-related lectins family

MIKYSKOVÁ, Michaela; Filip MELICHER a Michaela WIMMEROVÁ

Základní údaje

Originální název

Calocera viscosa lectin – a fungi member of the jacalin-related lectins family

Název česky

Calocera viscosa lektin - houbový zástupce lektinů podobných jacalinu

Autoři

MIKYSKOVÁ, Michaela (203 Česká republika, domácí); Filip MELICHER (703 Slovensko, domácí) a Michaela WIMMEROVÁ (203 Česká republika, garant, domácí)

Vydání

XXII. Meeting of biochemists and molecular biologists, 2023

Další údaje

Jazyk

angličtina

Typ výsledku

Prezentace na konferencích

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/23:00138791

Organizační jednotka

Přírodovědecká fakulta

ISBN

978-80-280-0409-5

Klíčová slova česky

lektiny; Calocera viscosa; krásnorůžek lepkavý; jacalinové lektiny; houby; sacharidy

Klíčová slova anglicky

lectins; Calocera viscosa; yellow stagshorn; jacalin-related lectins; fungi; saccharides

Změněno: 5. 2. 2025 17:03, Mgr. Michaela Mikysková

Anotace

V originále

The research is focused on the properties and structural analysis of the new lectin – saccharide binding protein – from the mushroom Calocera viscosa (CalVL). Mushroom lectins have been variously explored and studied for their potential in biomedicine and diagnostics. Based on their interactions with glycans, they exhibit mitogenic, antimicrobial, and antiproliferative effects.1 For the characterization of this lectin’s properties, various methods were employed. Inceptive predictions suggested that the CalVL, with its β-prism I fold, should belong to the family of jacalin related lectins (JRLs). JRLs may be briefly divided based on their preference of binding to D galactose and D-mannose.2 Molecular cloning and tests of gene expression were performed, and the CalVL was produced in the Escherichia coli expression system. Later, it was purified using affinity chromatography on mannose-agarose resin. Later it was shown by agglutination assays that the CalVL is able to agglutinate both yeast cells and human erythrocytes due to the interaction with their surface saccharides. Further analysis of the binding properties was inspected by the Glycan Array. Thermostability of the CalVL was determined by Differential Scanning Fluorimetry, homogeneity, and the oligomeric state of the protein by Dynamic Light Scattering and Analytical Ultracentrifugation. The CalVL’s ability to crystallize was tested in various screens using the vapor diffusion method, specifically the sitting drop technique. Data collection from obtained crystals was performed on synchrotron PETRA III and BESSY II in Hamburg and Berlin, respectively. The structure of the CalVL is in the process of solving but preliminary, there is a CalVL without any ligand, and with D mannose. The phase problem of the measured data was solved by molecular replacement, where the model of the CalVL predicted by the AlphaFold2 was utilized. These structures confirm the prediction of the CalVL being a member of the JRLs family.

Návaznosti

LM2023042, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CIISB - Česká infrastruktura pro integrativní strukturní biologii

Citovat

MIKYSKOVÁ, Michaela; Filip MELICHER a Michaela WIMMEROVÁ. Calocera viscosa lectin – a fungi member of the jacalin-related lectins family. In XXII. Meeting of biochemists and molecular biologists. 2023. ISBN 978-80-280-0409-5.

@proceedings{2474945,
   author = {Mikysková, Michaela and Melicher, Filip and Wimmerová, Michaela},
   booktitle = {XXII. Meeting of biochemists and molecular biologists},
   keywords = {lectins; Calocera viscosa; yellow stagshorn; jacalin-related lectins; fungi; saccharides},
   language = {eng},
   isbn = {978-80-280-0409-5},
   title = {Calocera viscosa lectin – a fungi member of the jacalin-related lectins family},
   year = {2023}
}

TY  - CONF
ID  - 2474945
AU  - Mikysková, Michaela - Melicher, Filip - Wimmerová, Michaela
PY  - 2023
TI  - Calocera viscosa lectin – a fungi member of the jacalin-related lectins family
SN  - 9788028004095
KW  - lectins
KW  - Calocera viscosa
KW  - yellow stagshorn
KW  - jacalin-related lectins
KW  - fungi
KW  - saccharides
N2  - The research is focused on the properties and structural analysis of the new lectin – saccharide binding protein – from the mushroom Calocera viscosa (CalVL). Mushroom lectins have been variously explored and studied for their potential in biomedicine and diagnostics. Based on their interactions with glycans, they exhibit mitogenic, antimicrobial, and antiproliferative effects.1 For the characterization of this lectin’s properties, various methods were employed. Inceptive predictions suggested that the CalVL, with its β-prism I fold, should belong to the family of jacalin related lectins (JRLs). JRLs may be briefly divided based on their preference of binding to D galactose and D-mannose.2 Molecular cloning and tests of gene expression were performed, and the CalVL was produced in the Escherichia coli expression system. Later, it was purified using affinity chromatography on mannose-agarose resin. Later it was shown by agglutination assays that the CalVL is able to agglutinate both yeast cells and human erythrocytes due to the interaction with their surface saccharides. Further analysis of the binding properties was inspected by the Glycan Array. Thermostability of the CalVL was determined by Differential Scanning Fluorimetry, homogeneity, and the oligomeric state of the protein by Dynamic Light Scattering and Analytical Ultracentrifugation. The CalVL’s ability to crystallize was tested in various screens using the vapor diffusion method, specifically the sitting drop technique. Data collection from obtained crystals was performed on synchrotron PETRA III and BESSY II in Hamburg and Berlin, respectively. The structure of the CalVL is in the process of solving but preliminary, there is a CalVL without any ligand, and with D mannose. The phase problem of the measured data was solved by molecular replacement, where the model of the CalVL predicted by the AlphaFold2 was utilized. These structures confirm the prediction of the CalVL being a member of the JRLs family.
ER  -

MIKYSKOVÁ, Michaela; Filip MELICHER a Michaela WIMMEROVÁ. Calocera viscosa lectin – a fungi member of the jacalin-related lectins family. In \textit{XXII. Meeting of biochemists and molecular biologists}. 2023. ISBN~978-80-280-0409-5.

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