High-performance adsorption chromatography of transfer ribonucleic acids and proteins on 2-microns spherical beads of hydroxyapatite. Influence of sodium chloride and magnesium ions on the resolution.
The influence of sodium chloride and magnesium chloride on the adsorption of tRNA and proteins on a high-performance liquid chromatographic column of 2-microns spherical hydroxyapatite beads was investigated. The resolution of 14C-labelled aminoacyl-tRNA isoacceptors was improved in the presence of sodium chloride. Inclusion of magnesium chloride in the buffers led to a separation of two tRNA species that could not be fractionated with or without sodium chloride in the eluting buffers (the original properties of the column were lost, however, and could not be regenerated by simply returning to magnesium chloride-free phosphate buffer). Also, the adsorption of some proteins was affected when salt was included in the buffers. For instance, the elution order of proteins could be changed by choosing an appropriate concentration of sodium chloride. This finding might be utilized to facilitate the purification of certain proteins.
Lindeberg
J
J
Institute of Biochemistry, University of Uppsala, Sweden.
Srichaiyo
T
T
Hjertén
S
S
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017886:Durapatite; D004926:Escherichia coli; D006882:Hydroxyapatites; D015636:Magnesium Chloride / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D011506:Proteins / Q000032:analysis* / Q000493:pharmacokinetics; D012343:RNA, Transfer / Q000032:analysis* / Q000187:drug effects / Q000493:pharmacokinetics; D012965:Sodium Chloride / Q000494:pharmacology
D006882:Hydroxyapatites; D011506:Proteins; D015636:Magnesium Chloride; D012965:Sodium Chloride; D011189:Potassium Chloride; D012343:RNA, Transfer; D017886:Durapatite