Open AccessCross‐linking of nucleic acids to proteins
Author(s) -
WOJTECH Eva,
BRIMACOMBE Richard,
HÄCKEL Andreas,
PROCHNOW Detlef,
FASOLD Hugo
Publication year - 1994
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1994.tb19055.x
Subject(s) - nucleic acid , ribosome , chemistry , biochemistry , protein subunit , escherichia coli , transfer rna , yield (engineering) , amino acid , nucleic acid analogue , rna , nucleic acid thermodynamics , gene , materials science , metallurgy
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 tRNALys 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.