Interaction of Galectin-1 with human NK cell receptors

• Thesis title in Czech: Interakce Galektinu-1 s receptory lidských NK buněk
• Thesis title in English: Interaction of Galectin-1 with human NK cell receptors
• Key words: NK buňky, CD69, Galektin-1, glykosylace, mikrotermoforézy
• English key words: NK cell, CD69, Galectin-1, glycosylation, microscale thermophoresis
• Academic year of topic announcement: 2016/2017
• Thesis type: diploma thesis
• Thesis language: angličtina
• Department: Department of Biochemistry (31-250)
• Supervisor: RNDr. Ondřej Vaněk, Ph.D.
• Author: hidden - assigned by the advisor, waiting for guarantor's approval
• Date of registration: 12.11.2016
• Date of assignment: 14.11.2016
• Date of electronic submission: 19.08.2019
• Date of proceeded defence: 03.09.2019
• Opponents: RNDr. Jiří Pavlíček, Ph.D.

Preliminary scope of work

Náplní práce bude rekombinantní exprese rozpustné formy receptoru CD69 (tzv. ranný aktivační antigen lidských lymfocytů) a jeho nedávno popsaných nových proteinových ligandů galektinu-1 a komplexu S100A8/S100A9 a studium jejich vzájemné interakce s cílem jejího strukturního popisu. Rekombinantní exprese proteinů v bakteriálním systému i v lidské buněčné linii HEK293, purifikace proteinů a jejich charakterizace pomocí analytické ultracentrifugace a rezonance povrchového plazmonu, krystalizace proteinů a řešení jejich struktury pomocí proteinové krystalografie.

Tato práce je součástí projektu "Harnessing soluble forms of NK cell receptors and their ligands for the generation of novel anticancer immunotherapeutics" podpořeného prestižní cenou Instruct R&D Pilot Project Award z evropského programu Instruct a také projektem Grantové agentury ČR.

Literatura:
The leukocyte activation receptor CD69 controls T cell differentiation through its interaction with galectin-1. de la Fuente H, Cruz-Adalia A, Martinez Del Hoyo G, Cibrián-Vera D, Bonay P, Pérez-Hernández D, Vázquez J, Navarro P, Gutierrez-Gallego R, Ramirez-Huesca M, Martín P, Sánchez-Madrid F. Mol Cell Biol. 2014;34(13):2479-87. doi: 10.1128/MCB.00348-14. PMID: 24752896.
Glycosylation-dependent interaction between CD69 and S100A8/S100A9 complex is required for regulatory T-cell differentiation. Lin CR, Wei TW, Tsai HY, Wu YT, Wu PY, Chen ST. FASEB J. 2015. pii: fj.15-273987. PMID: 26296369.

Preliminary scope of work in English

The aim of the thesis is recombinant expression of soluble form of receptor CD69 (so called "early activation antigen" of human lymphocytes) and its newly described protein ligands galectin-1 and S100A8/S100A9 complex and study of their mutual interaction with respect to its structural description. Recombinant protein expression in both bacterial and human HEK293 cell line expression systems, protein purification and characterization by analytical ultracentrifugation and surface plasmon resonance, protein crystallization and structure solution by protein crystallography.

This work is part of project "Harnessing soluble forms of NK cell receptors and their ligands for the generation of novel anticancer immunotherapeutics" supported by prestigious Instruct R&D Pilot Project Award from the European programe Instruct and also by project from Czech Science Foundation.

References:
The leukocyte activation receptor CD69 controls T cell differentiation through its interaction with galectin-1. de la Fuente H, Cruz-Adalia A, Martinez Del Hoyo G, Cibrián-Vera D, Bonay P, Pérez-Hernández D, Vázquez J, Navarro P, Gutierrez-Gallego R, Ramirez-Huesca M, Martín P, Sánchez-Madrid F. Mol Cell Biol. 2014;34(13):2479-87. doi: 10.1128/MCB.00348-14. PMID: 24752896.
Glycosylation-dependent interaction between CD69 and S100A8/S100A9 complex is required for regulatory T-cell differentiation. Lin CR, Wei TW, Tsai HY, Wu YT, Wu PY, Chen ST. FASEB J. 2015. pii: fj.15-273987. PMID: 26296369.