Analysis of protein interactors of cytosolic seryl-tRNA synthetase from plant Arabidopsis thaliana

Zanki, Vladimir (2016) Analysis of protein interactors of cytosolic seryl-tRNA synthetase from plant Arabidopsis thaliana. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

Seryl-tRNA synthetases (SerRS) are enzymes responsible for covalent attachment of cognate tRNA with amino acid serine thus creating a foundation for accurate protein biosynthesis. In addition to this canonical role, SerRSs participate in other noncanonical pathways either independently or via interactions with other cellular proteins. In this thesis, recently identified potential interaction partners of the cytosolic SerRS from plant Arabidopsis thaliana, glutathione-S-transferase phi 2 (GSTF2) and protein of unknown function (PNF) were prepared for biophysical analysis, while gene encoding ribosomal protein L9 was successfully cloned in plasmid vectors. Furthermore, in order to determine interaction surface between cytosolic protein SerRS and recently identified interaction partner, protein BEN1 involved in brassinosteroid hormone metabolism, truncated variants of these proteins were designed. Microthermophoresis measurements have shown that cytosolic SerRS interacts with the metabolic protein BEN1 using its central catalytic domain, while protein BEN1 interacts with SerRS using its N-terminal unstructured hydrophilic extension. In addition, significant improvement in yield and stability of the catalytic domain of cytosolic SerRS using the GST-tag was also observed.

Item Type: Thesis (Diploma thesis)
Keywords: recombinant DNA technology, protein purification, protein interactions, microscale thermophoresis, seryl-tRNA synthetase, Arabidopsis thaliana
Supervisor: Rokov Plavec, Jasmina and Kekez, Mario
Date: 2016
Number of Pages: 115
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 21 Oct 2016 09:18
Last Modified: 21 Oct 2016 12:00
URI: http://digre.pmf.unizg.hr/id/eprint/5206

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