Total reconstitution of active small ribosomal subunits of the extreme halophilic archaeon Haloferax mediterranei.
The small ribosomal subunit of the halophilic archaeon Haloferax mediterranei has been reconstituted from its dissociated rRNA and protein components. Efficient reconstitution of particles, fully active in poly(U)-dependent polyphenylalanine synthesis, occurs after 2 h of incubation at 36 degrees C in the presence of 1.5 M of (NH4)2SO4, 100 mM of MgAc2, 20 mM Tris-HCl (pH 8.2) and 6 mM 2-mercaptoethanol. Important differences in the optimal ionic conditions for the reconstitution of the 30S and the 50S ribosomal subunits from Haloferax mediterranei have been found. K+ and NH4+ ions have differing abilities to promote the reconstitution of the particles. The assembly of 30S ribosomal subunits of H. mediterranei has a higher tolerance to ionic strength than the assembly of the 50S subunits and it is independent of the Mg2+ concentration present in the system.
Sánchez
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ME
Centro de Biología Molecular, C.S.I.C.-U.A.M., Cantoblanco, Madrid, Spain.
Londei
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P
Amils
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R
D006216:Halobacteriaceae / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D010452:Peptide Biosynthesis; D010455:Peptides; D011188:Potassium / Q000494:pharmacology; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D012335:RNA, Ribosomal / Q000378:metabolism; D012269:Ribosomal Proteins / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism*; D012964:Sodium / Q000494:pharmacology; D013696:Temperature; D014461:Ultracentrifugation
D010455:Peptides; D000644:Quaternary Ammonium Compounds; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; C012001:polyphenylalanine; D012964:Sodium; D008274:Magnesium; D011188:Potassium