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Preliminary crystallographic study of Streptomyces coelicolor single-stranded DNA-binding protein

Štefanić, Zoran; Vujaklija, Dušica; Andrišić, Luka; Mikleušević, Goran; Andrejašič, Miha; Turk, Dušan; Luić, Marija (2007) Preliminary crystallographic study of Streptomyces coelicolor single-stranded DNA-binding protein. Croatica Chemica Acta, 80 (1). pp. 35-39. ISSN 0011-1643

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Abstract

Single-stranded DNA-binding proteins (SSBs) play a crucial role in DNA processing such as replication, repair and recombination in all organisms, from bacteria to human. Streptomyces coelicolor ssb gene was overexpressed in a heterologous host, Escherichia coli NM522. 15 mg of purified protein from 1 dm(3) of culture was obtained in one-step procedure applying Ni2+ chelating chromatography. Among bacterial SSBs with the solved crystal structure, the S. coelicolor SSB displayed significant sequence similarity with those from Mycobacterium tuberculosis and Mycobacterium smegmatis, slow growing bacteria with a high GC content. Moreover, conserved amino acid region that forms additional B strand in mycobacterial SSBs was also found in S. coelicolor SSB. The full-length protein readily crystallises in space group 1222 or I2(1)2(1)2(1) with unit-cell parameters a = 100.8, b = 102.1, c = 164.2 angstrom. The asymmetric unit is expected to contain four monomers with solvent content of 52-55 %.

Item Type: Article
Uncontrolled Keywords: single-stranded DNA-binding protein; SSB purification; Streptomyces coelicolor; crystallisation; crystal-structure; diffraction data; crystallization; purification; variability; tyrosine; quality
Subjects: NATURAL SCIENCES > Chemistry
Divisions: Division of Molecular Biology
Division of Molecular Medicine
Division of Physical Chemistry
Projects:
Project titleProject leaderProject codeProject type
Protein-ligand međudjelovanja na atomnoj razini[77430] Marija Luić098-1191344-2943MZOS
Temeljna molekularno-biološka istraživanja streptomiceta[116760] Dušica Vujaklija098-0982913-2877MZOS
Depositing User: Marija Luić
Date Deposited: 25 Oct 2013 15:05
Last Modified: 14 Feb 2014 15:11
URI: http://fulir.irb.hr/id/eprint/849

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