Immunoblotting analysis of protein-protein crosslinks within the 50S ribosomal subunit of Escherichia coli. A study using dimethylsuberimidate as crosslinking reagent.
50S ribosomal subunits of Escherichia coli have been crosslinked with the bifunctional imidoester dimethyl-suberimidate and the protein-protein crosslinks have been analyzed by immunoblotting, using antisera specific for the individual ribosomal proteins of the large ribosomal subunit. Crosslinked protein pairs which occurred in yields higher than 5% have been unambiguously identified. Thus 13 crosslinks have been identified, namely L1-L33, L5-L7/12, L6-L19, L7/12-L10, L7/12-L11, L9-L28, L10-L11, L13-L20, L16-L27, L17-L32, L18-L22, L19-L25 and L27-L33. These data, together with the results which we will be presenting elsewhere, contribute considerably to our knowledge of the protein topography of the 50S ribosomal proteins as determined by immunoelectron microscopy. We can now propose the approximate locations of ten proteins that have not previously been localized.
Redl
B
B
Institut für Mikrobiologie, Medizinische Fakultät der Universität, Innsbruck, Austria.
Walleczek
J
J
Stöffler-Meilicke
M
M
Stöffler
G
G
D004120:Dimethyl Suberimidate; D004926:Escherichia coli / Q000032:analysis*; D007096:Imidoesters; D015151:Immunoblotting / Q000379:methods; D008961:Models, Structural; D008970:Molecular Weight; D012269:Ribosomal Proteins / Q000032:analysis*; D012270:Ribosomes / Q000032:analysis* / Q000648:ultrastructure
D007096:Imidoesters; D012269:Ribosomal Proteins; D004120:Dimethyl Suberimidate