Expression, purification, crystallization and preliminary diffraction analysis of RNase P protein from Thermotoga maritima.
Ribonuclease P (RNase P), the ubiquitous endonuclease that catalyzes maturation of the 5'-end of tRNA in bacteria, is a ribonucleoprotein particle composed of one large RNA and one small protein. Two major structural types of bacterial RNase P RNA have been identified by phylogenetic comparative analysis: the A (ancestral) and B (Bacillus) types. The RNase P protein from Thermotoga maritima, a hyperthermophilic bacterium with an A-type RNase P RNA, has been expressed in Escherichia coli. A purification strategy was developed to obtain a protein preparation suitable for crystallization. Protein crystals suitable for diffraction studies were obtained and characterized.
Krivenko
Angelika A
AA
Department MCD Biology, University of Colorado, Boulder 80309, USA.
Kazantsev
Alexei V
AV
Adamidi
Catherine
C
Harrington
Daniel J
DJ
Pace
Norman R
NR
D018360:Crystallography, X-Ray / Q000379:methods*; D004722:Endoribonucleases / Q000737:chemistry* / Q000302:isolation & purification*; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D016337:RNA, Catalytic / Q000737:chemistry* / Q000302:isolation & purification*; D043262:Ribonuclease P; D020124:Thermotoga maritima / Q000201:enzymology*
D029968:Escherichia coli Proteins; D016337:RNA, Catalytic; D004722:Endoribonucleases; D043262:Ribonuclease P; C470865:ribonuclease P, E coli