Contamination of ribosome inactivating proteins with ribonucleases, separated by affinity chromatography on red sepharose.
Three preparations of type 1 ribosome inactivating proteins (RIPs), namely, agrostin, saporin, and luffin, were subjected to affinity chromatography on Red Sepharose and eluted with a linear concentration gradient of NaCl in 10 mM Tris-HCl buffer (pH 7.4). The eluate was assayed for ability to inhibit translation in a cell-free rabbit reticulocyte lysate system which measures RIP activity, and for ability to hydrolyze yeast transfer RNA which measures RNase activity. It was found that, in all three RIP preparations, the peak of RIP activity, which coincided with the peak of absorbance at 280 nm, was eluted earlier than the peak of RNase activity. It appears that RNase is a possible contaminant of ribosome inactivating protein preparations and that this contamination can be minimized by using Red Sepharose.
Wang
H X
HX
Department of Microbiology, China Agricultural University, Beijing, P. R. China.
Ng
T B
TB
Cheng
C H K
CH
Fong
W P
WP
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D000922:Immunotoxins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D009699:N-Glycosyl Hydrolases / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D010944:Plants / Q000737:chemistry; D014176:Protein Biosynthesis / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012343:RNA, Transfer / Q000378:metabolism; D011817:Rabbits; D012260:Ribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins; D012639:Seeds / Q000737:chemistry; D012685:Sepharose / Q000031:analogs & derivatives
D000922:Immunotoxins; D010940:Plant Proteins; D011500:Protein Synthesis Inhibitors; D054789:Ribosome Inactivating Proteins, Type 1; C459705:agrostin protein, Agrostemma; C063258:luffin-a protein, Luffa cylindrica; D012685:Sepharose; D012343:RNA, Transfer; D012260:Ribonucleases; D009699:N-Glycosyl Hydrolases; D000076985:Saporins