Cross-linking of nucleic acids to proteins. Modified poly(A) as mRNA for Escherichia coli ribosomes.
Poly(adenylic acid) was modified by methylchlorotetrolic ester in a reproducible and defined content of the derivatized bases. The nucleic acid derivative is protein reactive and was coupled to 70S ribosomes from Escherichia coli, in order to identify proteins along the mRNA pathway. The binding of the label becomes specific under the direction of tRNA(Lys) and is then almost exclusively located on the small subunit. The proteins S1, S12, S18 and S21 were labeled, as shown by an antibody assay. The yield of the affinity label was 5.4%, as calculated from the labeled nucleic acid. This compares favourably with the yields from photolabile compounds.
Wojtech
E
E
Institut für Biochemie, J. W. Goethe-Universität Frankfurt, Germany.
Brimacombe
R
R
Häckel
A
A
Prochnow
D
D
Fasold
H
H
D000345:Affinity Labels; D000480:Alkynes / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry; D011061:Poly A / Q000737:chemistry*; D012333:RNA, Messenger / Q000737:chemistry*; D012269:Ribosomal Proteins / Q000737:chemistry; D012270:Ribosomes / Q000737:chemistry*
D000345:Affinity Labels; D000480:Alkynes; D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D012333:RNA, Messenger; D012269:Ribosomal Proteins; D011061:Poly A; C051334:methyl chlorotetrolate