Journal Article

PUBDB-2016-05653

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png 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

Grinkevich, P. ; Iermak, I. ; Luedtke, N. A. ; Mesters, J. R. ; Ettrich, R. (Corresponding author) ; Ludwig, J.

2016
IUCr

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Please use a persistent id in citations: doi:10.1107/S2053230X16011626  doi:10.3204/PUBDB-2016-05653

Abstract: 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 Å, $\alpha$ = $\beta$ = $\gamma$ = 90.00° and two molecules in the asymmetric unit (V$_M$ = 2.55 Å$^3$ Da$^{-1}$, solvent content 50.47%). X-ray diffraction data were collected to a resolution of 2.45 Å.

Note: (c) International Union of Crystallography

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Contributing Institute(s):

1. EMBL-User (EMBL-User)

Research Program(s):

1. 6G3 - PETRA III (POF3-622) (POF3-622)
2. BIOSTRUCT-X - Transnational access and enhancement of integrated Biological Structure determination at synchrotron X-ray radiation facilities (283570) (283570)

Experiment(s):

1. PETRA Beamline P13 (PETRA III)

Appears in the scientific report 2016

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Database coverage:
; ; BIOSIS Previews ; Ebsco Academic Search ; IF < 5 ; JCR ; NCBI Molecular Biology Database ; SCOPUS ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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