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 title Project leader Project code Project type Protein-ligand međudjelovanja na atomnoj razini [77430] Marija Luić 098-1191344-2943 MZOS Temeljna molekularno-biološka istraživanja streptomiceta [116760] Dušica Vujaklija 098-0982913-2877 MZOS
Depositing User:	Marija Luić
Date Deposited:	25 Oct 2013 15:05
URI:	http://fulir.irb.hr/id/eprint/849

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