Isolation of ribosomal subunits containing intact rRNA from human placenta: estimation of functional activity of 80S ribosomes.
We have elaborated a method for the isolation of ribosomal subunits from fresh unfrozen human placenta containing intact rRNA and a complete set of ribosomal proteins. Activity of 80S ribosomes obtained by reassociation of 40S and 60S subunits in nonenzymatic poly(U)-dependent binding of Phe-tRNA(Phe) was equal to 80% (above 1.5 mol [14C]Phe-tRNA(Phe) is coupled to 1 mol of ribosomes). The activity of 80S ribosomes in poly(U)-directed synthesis of polyphenylalanine was tested in a polysome-free protein-synthesizing system from rabbit reticulocytes. About 100 mol of phenylalanine residue was polymerized by a mole of ribosomes at a rate of 0.83 residues per minute in this system (2 h, 37 degrees C).
Matasova
N B
NB
Institute of Bioorganic Chemistry, Siberian Division of the USSR Academy of Sciences, Novosibirsk.
Myltseva
S V
SV
Zenkova
M A
MA
Graifer
D M
DM
Vladimirov
S N
SN
Karpova
G G
GG
D000818:Animals; D001665:Binding Sites; D002458:Cell Fractionation; D002474:Cell-Free System; D006801:Humans; D010452:Peptide Biosynthesis; D010455:Peptides; D010920:Placenta / Q000737:chemistry*; D011072:Poly U; D012335:RNA, Ribosomal / Q000302:isolation & purification*; D012360:RNA, Transfer, Phe / Q000737:chemistry; D011817:Rabbits; D012156:Reticulocytes; D012269:Ribosomal Proteins / Q000302:isolation & purification; D012270:Ribosomes / Q000737:chemistry* / Q000502:physiology
D010455:Peptides; D012335:RNA, Ribosomal; D012360:RNA, Transfer, Phe; D012269:Ribosomal Proteins; D011072:Poly U; C012001:polyphenylalanine