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

Autori
Štefanić, Zoran ; Vujaklija, Dušica ; Andrišić, Luka ; Mikleušević, Goran ; Andrejašič, Miha ; Turk, Dušan ; Luić, Marija

Izvornik
Croatica chemica acta (0011-1643) 80 (2007), 1; 35-39

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
single-stranded DNA-binding protein ; SSB purification ; Streptomyces coelicolor ; crystallisation
(Single-stranded DNA-binding protein ; SSB purification ; Streptomyces coelicolor ; crystallisation)

Sažetak
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 dm3 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 β strand in mycobacterial SSBs was also found in S. coelicolor SSB. The full-length protein readily crystallises in space group I222 or I212121 with unit-cell parameters a = 100.8, b = 102.1, c = 164.2 Å ; . The asymmetric unit is expected to contain four monomers with solvent content of 52-55 %.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija

POVEZANOST RADA