Simple and rapid chromatographic method for the separation of major classes of ribosomal ribomononucleotides.
We have described a simple and rapid chromatographic method for the analytical and preparative separation of major types of ribosomal ribomononucleotides with Dowex 1-X10 (HCOO-, 37-74 microns) and Dowex 2-X10 (HCOO-, 37-74 microns) columns, by desorption with formiate solutions in 1-2 h. The separation has been achieved for Cp, Ap, Up and Gp, while a mixture of 2'-, and 3'-nucleoside phosphates desorbs as a single peak; with both resins, a successful separation was achieved with a load from 25 micrograms to 1 mg of ribomononucleotide mixture per ml of packed resin. A complete separation was achieved with Dowex 1, while the separation with Dowex 2 resin was even better. The resins cannot separate unusual nucleosides; therefore, our method is suitable for studies of ribonucleic acids with a low content of unusual nucleosides. Our method has been applied for the quantitative determination of the ribomononucleotide composition of 18S and 28S rRNAs, isolated from mammalian tissues: rat liver, mouse kidney and Ehrlich ascites cells. Dowex 1 and Dowex 2 resins afforded similar or identical ribomononucleotide compositions in all cases; analytical data were in agreement with the literature data. Our method is competitive, in several respects, with modern HPLC techniques for the separation of ribomononucleotides.
Popović
D A
DA
Department of Biochemistry, Medical Faculty, Novi Sad, Yugoslavia.
Leskovac
V
V
D000818:Animals; D000837:Anion Exchange Resins / Q000378:metabolism; D002286:Carcinoma, Ehrlich Tumor / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D006868:Hydrolysis; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D051379:Mice; D012335:RNA, Ribosomal / Q000032:analysis / Q000737:chemistry*; D012337:RNA, Ribosomal, 18S / Q000737:chemistry; D012339:RNA, Ribosomal, 28S / Q000737:chemistry; D051381:Rats; D012117:Resins, Synthetic; D012265:Ribonucleotides / Q000032:analysis / Q000302:isolation & purification*; D012270:Ribosomes / Q000737:chemistry*
D000837:Anion Exchange Resins; D012335:RNA, Ribosomal; D012337:RNA, Ribosomal, 18S; D012339:RNA, Ribosomal, 28S; D012117:Resins, Synthetic; D012265:Ribonucleotides; C027972:Dowex