High-performance liquid chromatographic purification of a ribonucleoprotein complex and its protein component.
This paper describes the rapid purification by high-performance liquid chromatographic techniques of milligram quantities of the 7 S ribonucleoprotein complex (RNP 7 S) and its protein component from tench (Tinca tinca) oocyte extracts. High-performance gel permeation chromatography (with a TSK 3000SW column) was found to be unacceptable because of multiple contaminants which coelute with RNP 7 S. In contrast, semipreparative high-performance DEAE ion-exchange chromatography was found to give an excellent separation of the 7 S complex which could be directly adapted to a preparative scale providing rapid purification (less than 1 h) of milligram quantities of the complex. Agarose electrophoresis followed by specific staining of protein and nucleic acid was found to be a convenient and rapid means of evaluating the purification. Finally, reverse-phase high-performance liquid chromatography was found suitable for the purification of the protein component of the 7 S complex.
Monteilhet
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Aggerbeck
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D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange; D004587:Electrophoresis, Agar Gel; D005260:Female; D005399:Fishes; D008970:Molecular Weight; D009698:Nucleoproteins / Q000302:isolation & purification*; D009865:Oocytes / Q000032:analysis; D012261:Ribonucleoproteins / Q000302:isolation & purification*
D009698:Nucleoproteins; D012261:Ribonucleoproteins