Crystallization and structure determination of delta-subunit of RNApolymerase

DEMO, Gabriel, Veronika PAPOUŠKOVÁ, Hana ŠANDEROVÁ, Lukáš ŽÍDEK and Michaela WIMMEROVÁ. Crystallization and structure determination of delta-subunit of RNApolymerase. In Higher European Research Course for Users of Large Experimental Systems (HERCULES). 2011.

Basic information

• Original name: Crystallization and structure determination of delta-subunit of RNApolymerase
• Name in Czech: Krystalizace a určení struktury delta-podjednotky RNA polymerázy
• Authors: DEMO, Gabriel (703 Slovakia, belonging to the institution), Veronika PAPOUŠKOVÁ (203 Czech Republic, belonging to the institution), Hana ŠANDEROVÁ (203 Czech Republic), Lukáš ŽÍDEK (203 Czech Republic, belonging to the institution) and Michaela WIMMEROVÁ (203 Czech Republic, guarantor, belonging to the institution).
• Edition: Higher European Research Course for Users of Large Experimental Systems (HERCULES), 2011.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• RIV identification code: RIV/00216224:14740/11:00049774
• Organization unit: Central European Institute of Technology
• Keywords (in Czech): delta-podjednotka RNA polymeráza krystalizace struktura
• Keywords in English: delta-subunit RNA polymerase crystallization structure
• Tags: rivok

Abstract

The structure of N-terminal domain was determined by NMR. It consists mainly of three alpha-helixes and one short beta-sheet, yet the N-terminal part remains unstructured. The cause of this flexibility is probably the His6-Tag attached at the N-terminus.The N-terminal domain of delta subunit was conquested to high-throughput screening (sitting drop), where several crystallization conditions were found.Higher quality crystals were obtained after 7-9 days with an average size 150 x 100 micrometers.Diffraction data were collected at ESRF Grenoble, ID-23. The resolution of the structure was 3.5 Angstrom. The data were processed by MOSFLM and the determination of the structure was done by molecular replacement with NMR structure used as a model.

Abstract (in Czech)

Struktura N-terminální domény byla určená NMR. Její struktura je tvořena 3 alfa-helixy a jedním krátkym beta-listem. Část domény zůstáva neuspořádaná, pravděpodobně díky flexibilnímu konci His6-Tag. Přistoupilo se ke krystalizaci N-terminální domény, kde byli najdeny krystalizační podmínky pro získání krystalu. Krystaly byli získany po 7-9 dnech s průměrnou velikostí 150 x 100 mikrometru. Difrakční data byli naměřeny na ESRF v Grenobli na ID-23. Rozlišení dat bylo 3.5 Angstrom. Data byli spracovány programem MOSFLM a struktura delta podjednotky byla určená molekulárním nahrazením za použití NMR struktury jako modelu.

Links

• GA204/09/0583, research and development project: Name: Delta podjednotka RNA polymerázy z Gram pozitivních bakterií (Acronym: DELTA)

Investor: Czech Science Foundation

• GD301/09/H004, research and development project: Name: Molekulární a strukturní biologie vybraných cytostatik. Od mechanistických studií k chemoterapii rakoviny

Investor: Czech Science Foundation

• MSM0021622413, plan (intention): Name: Proteiny v metabolismu a při interakci organismů s prostředím

Investor: Ministry of Education, Youth and Sports of the CR, Proteins in metabolism and interaction of organisms with the environment