pHluorin-assisted expression, purification, crystallization and X-ray diffraction data analysis of the C-terminal domain of the HsdR subunit of the Escherichia coli type I restriction-modification system EcoR124I

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F16%3A00469849" target="_blank" >RIV/61388971:_____/16:00469849 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/60076658:12310/16:43891107 RIV/60076658:12520/16:43891107

Výsledek na webu

<a href="http://dx.doi.org/10.1107/S2053230X16011626" target="_blank" >http://dx.doi.org/10.1107/S2053230X16011626</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1107/S2053230X16011626" target="_blank" >10.1107/S2053230X16011626</a>

Jazyk výsledku

angličtina

Název v původním jazyce

pHluorin-assisted expression, purification, crystallization and X-ray diffraction data analysis of the C-terminal domain of the HsdR subunit of the Escherichia coli type I restriction-modification system EcoR124I

Popis výsledku v původním jazyce

The HsdR subunit of the type I restriction-modification system EcoR124I is responsible for the translocation as well as the restriction activity of the whole complex consisting of the HsdR, HsdM and HsdS subunits, and while crystal structures are available for the wild type and several mutants, the C-terminal domain comprising approximately 150 residues was not resolved in any of these structures. Here, three fusion constructs with the GFP variant pHluorin developed to overexpress, purify and crystallize the C-terminal domain of HsdR are reported. The shortest of the three encompassed HsdR residues 887-1038 and yielded crystals that belonged to the orthorhombic space group C222(1), with unit-cell parameters a = 83.42, b = 176.58, c = 126.03 angstrom, alpha = beta = gamma = 90.00 degrees and two molecules in the asymmetric unit (V-M = 2.55 angstrom(3) Da(-1), solvent content 50.47%). X-ray diffraction data were collected to a resolution of 2.45 angstrom.

Název v anglickém jazyce

pHluorin-assisted expression, purification, crystallization and X-ray diffraction data analysis of the C-terminal domain of the HsdR subunit of the Escherichia coli type I restriction-modification system EcoR124I

Popis výsledku anglicky

The HsdR subunit of the type I restriction-modification system EcoR124I is responsible for the translocation as well as the restriction activity of the whole complex consisting of the HsdR, HsdM and HsdS subunits, and while crystal structures are available for the wild type and several mutants, the C-terminal domain comprising approximately 150 residues was not resolved in any of these structures. Here, three fusion constructs with the GFP variant pHluorin developed to overexpress, purify and crystallize the C-terminal domain of HsdR are reported. The shortest of the three encompassed HsdR residues 887-1038 and yielded crystals that belonged to the orthorhombic space group C222(1), with unit-cell parameters a = 83.42, b = 176.58, c = 126.03 angstrom, alpha = beta = gamma = 90.00 degrees and two molecules in the asymmetric unit (V-M = 2.55 angstrom(3) Da(-1), solvent content 50.47%). X-ray diffraction data were collected to a resolution of 2.45 angstrom.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EE - Mikrobiologie, virologie

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2016

Kód důvěrnosti údajů

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

Název periodika

ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS

ISSN

2053-230X

e-ISSN

—

Svazek periodika

72

Číslo periodika v rámci svazku

9

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

5

Strana od-do

672-676

Kód UT WoS článku

000384167800003

EID výsledku v databázi Scopus

2-s2.0-84986269515