Plasmid preparation: Optimization of 17-β-hydroxysteroid dehydrogenase 1 (HSD17B1) recombinant expression

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F23%3A10477837" target="_blank" >RIV/00216208:11310/23:10477837 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Plasmid preparation: Optimization of 17-β-hydroxysteroid dehydrogenase 1 (HSD17B1) recombinant expression

Popis výsledku v původním jazyce

Breast cancer, the most common cancer among women in western populations with a 10% lifetime risk, is largely driven by estrogen. Estradiol (E2), the most potent estrogen, promotes breast cancer growth. 17-β-hydroxysteroid dehydrogenases (HSD17B) catalyze E2 formation. HSD17B1, highly expressed in placenta, ovaries, and breast cancer tissue, is particularly prominent in aggressive hormone-dependent cancers, correlating with poor prognosis and short recurrence times. Inhibiting HSD17B1 presents a potential avenue for treating estrogen receptor-positive breast cancer.Attempts to express HSD17B1 in E. coli yielded minimal soluble protein and activity, limiting inhibitor screening via co-crystallization or enzyme assays. Expression in HEK293T cells resulted in low yields (approx. 2 mg/liter) of active protein from transient transfection. However, this expression level and purity are insufficient for cost-effective inhibitor screening. Additionally, wild-type enzyme activity diminishes upon long-term storage, necessitating the search for more stable protein mutants with higher yields, while preserving the active site and homodimer interface, as explored through the PROSS benchmark.

Název v anglickém jazyce

Plasmid preparation: Optimization of 17-β-hydroxysteroid dehydrogenase 1 (HSD17B1) recombinant expression

Popis výsledku anglicky

Breast cancer, the most common cancer among women in western populations with a 10% lifetime risk, is largely driven by estrogen. Estradiol (E2), the most potent estrogen, promotes breast cancer growth. 17-β-hydroxysteroid dehydrogenases (HSD17B) catalyze E2 formation. HSD17B1, highly expressed in placenta, ovaries, and breast cancer tissue, is particularly prominent in aggressive hormone-dependent cancers, correlating with poor prognosis and short recurrence times. Inhibiting HSD17B1 presents a potential avenue for treating estrogen receptor-positive breast cancer.Attempts to express HSD17B1 in E. coli yielded minimal soluble protein and activity, limiting inhibitor screening via co-crystallization or enzyme assays. Expression in HEK293T cells resulted in low yields (approx. 2 mg/liter) of active protein from transient transfection. However, this expression level and purity are insufficient for cost-effective inhibitor screening. Additionally, wild-type enzyme activity diminishes upon long-term storage, necessitating the search for more stable protein mutants with higher yields, while preserving the active site and homodimer interface, as explored through the PROSS benchmark.

Druh

V_souhrn - Souhrnná výzkumná zpráva

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

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Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2023

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Počet stran výsledku

8

Místo vydání

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Název nakladatele resp. objednatele

IOCB TECH s.r.o., Flemingovo náměstí 542/2, 160 00 Praha 6, Česká republika

Verze

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