diploma thesis
Nikola Janakievski (Avtor), Aljaž Gaber (Mentor), Maria dos Anjos Macedo (Komentor)

Povzetek

Gla-rich protein (GRP) is a novel protein connected to multiple pathological disorders plaguing humanity in modern times, including chronic kidney disease, atherosclerosis, osteoarthritis, and cancer. It gets its name from the high number of γ-carboxylated glutamate residues, most of any known protein, this enables it to have a very high affinity for calcium ions, making it a potential inhibitor of ectopic calcification in soft tissue. Except for the fact that this protein contains intrinsically disordered domains, not much else is known about the structure of GRP. NMR spectroscopy is the perfect structure determination technique to tackle this problem, specializing in the structural determination of small proteins and more recently for intrinsically disordered proteins. We expressed the human GRP variant in transformed E. coli cells and labelled it with isotopes 13C and 15N. After purification, we prepared samples for liquid-state NMR spectroscopy and explored the most practical NMR experiments for structure determination. To conclude, we found the optimal experimental conditions for NMR sample preparation of human GRP allowing for structural studies to be performed. We tested out different 15N and 13C direct detection experiments at different acquisition temperatures. Our preliminary data is promising and with the acquisition of other types of multidimensional NMR spectra, and after resonance assignment, structural information can be obtained for human GRP structural features elucidation.

Ključne besede

NMR spectroscopy;ectopic calcification;human GRP;expression;IDPs;

Podatki

Jezik: Angleški jezik
Leto izida:
Tipologija: 2.11 - Diplomsko delo
Organizacija: UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik: [N. Janakievski]
UDK: 577.112(043.2)
COBISS: 138709763 Povezava se bo odprla v novem oknu
Št. ogledov: 41
Št. prenosov: 10
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Slovenski jezik
Sekundarni naslov: Priprava in strukturna karakterizacija inhibitorja žilne kalcinacije, človeškega s karboksiglutaminsko kislino bogatega proteina (GRP)
Sekundarni povzetek: Gla-rich protein (GRP) is a novel protein connected to multiple pathological disorders plaguing humanity in modern times, including chronic kidney disease, atherosclerosis, osteoarthritis, and cancer. It gets its name from the high number of γ-carboxylated glutamate residues, most of any known protein, this enables it to have a very high affinity for calcium ions, making it a potential inhibitor of ectopic calcification in soft tissue. Except for the fact that this protein contains intrinsically disordered domains, not much else is known about the structure of GRP. NMR spectroscopy is the perfect structure determination technique to tackle this problem, specializing in the structural determination of small proteins and more recently for intrinsically disordered proteins. We expressed the human GRP variant in transformed E. coli cells and labelled it with isotopes 13C and 15N. After purification, we prepared samples for liquid-state NMR spectroscopy and explored the most practical NMR experiments for structure determination. To conclude, we found the optimal experimental conditions for NMR sample preparation of human GRP allowing for structural studies to be performed. We tested out different 15N and 13C direct detection experiments at different acquisition temperatures. Our preliminary data is promising and with the acquisition of other types of multidimensional NMR spectra, and after resonance assignment, structural information can be obtained for human GRP structural features elucidation.
Sekundarne ključne besede: človeški GRP;IDP;ekspresija;ektopična kalcifikacija;NMR spektroskopija;diplomska dela;Beljakovine;Univerzitetna in visokošolska dela;
Vrsta dela (COBISS): Diplomsko delo/naloga
Študijski program: 1000371
Konec prepovedi (OpenAIRE): 1970-01-01
Komentar na gradivo: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, UNI Biokemija
Strani: 36 str.
ID: 17389064