title: Removal of protein S1 from Escherichia coli ribosomes without the use of affinity chromatography.
abstract: The paper reports an inexpensive and efficient procedure for the removal of protein S1 from E. coli ribosomes. It comprises incubation of ribosomes in a pyrimidine polyribonucleotide solution followed by centrifugation of the sample through a sucrose cushion. To avoid co-sedimentation of the S1-bound polypyrimidine with the ribosomes, its length should not exceed several hundred nucleotides. Unlike popular affinity chromatography through a poly(U) Sepharose or poly(U) cellulose column, the method tolerates limited polyribonucleotide degradation by eventual traces of ribonucleases, and can readily be incorporated into standard protocols for the isolation of ribosomes by centrifugation.
authors.affiliation: Institute of Protein Research of the Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia.
authors.forename: Marianna D
authors.initials: MD
authors.lastname: Baranovskaya
authors.affiliation: Institute of Protein Research of the Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia.
authors.forename: Victor I
authors.initials: VI
authors.lastname: Ugarov
authors.affiliation: Institute of Protein Research of the Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia.
authors.forename: Helena V
authors.initials: HV
authors.lastname: Chetverina
authors.affiliation: Institute of Protein Research of the Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia. Electronic address: alexch@vega.protres.ru.
authors.forename: Alexander B
authors.initials: AB
authors.lastname: Chetverin
mesh_terms: D002499:Centrifugation, Density Gradient / Q000379:methods; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000302:isolation & purification*; D011072:Poly U / Q000737:chemistry; D012269:Ribosomal Proteins / Q000302:isolation & purification*; D012270:Ribosomes / Q000737:chemistry*; D013395:Sucrose / Q000031:analogs & derivatives / Q000737:chemistry
chemical_list: D029968:Escherichia coli Proteins; D012269:Ribosomal Proteins; C532344:Sucrocide; C544829:ribosomal protein S1, E coli; D011072:Poly U; D013395:Sucrose
keywords: Centrifugation; E. coli ribosomes; Poly(C); Poly(U); Protein S1