Placement of dinitrophenyl-modified ribosomal proteins in totally reconstituted Escherichia coli 30 S subunits. Localization of proteins S6, S13, S16, and S18 by immune electron microscopy.
Purified Escherichia coli ribosomal proteins S6, S13, S16, and S18 were dinitrophenylated at their amino termini and/or at one or more internal lysine residues. Each dinitrophenyl protein was then separately incorporated into reconstituted small ribosomal subunits. Modified proteins were localized on the 30 S subunit surface by electron microscopy of reconstituted subunits complexed with antibodies to dinitrophenol (DNP). DNP protein S13 was placed on the subunit head above the platform and on the surface that faces the large subunit. DNP-S18 was localized to the subunit platform below the tip and in a region associated with binding to 50 S subunits. DNP proteins S6 and S16 were both localized near the junction of the subunit body and platform; DNP-S6 was available to antibody in 70 S ribosomes and was placed on the cytoplasm-facing side of the subunit in an area that overlaps the platform and body of the particle. DNP-S16 in 70 S ribosomes was not bound by antibody. It was localized to the 30 S body near its junction with the platform and on the surface facing the 50 S particle. The results complement and clarify data obtained using other approaches.
Montesano-Roditis
L
L
Department of Biological Chemistry, UCLA School of Medicine 90024-1737.
McWilliams
R
R
Glitz
D G
DG
Olah
T V
TV
Perrault
A R
AR
Cooperman
B S
BS
D000936:Antigen-Antibody Complex; D002851:Chromatography, High Pressure Liquid; D004139:Dinitrofluorobenzene / Q000276:immunology / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D007074:Immunoglobulin G; D016253:Microscopy, Immunoelectron; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins / Q000032:analysis* / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure
D000936:Antigen-Antibody Complex; D029968:Escherichia coli Proteins; D007074:Immunoglobulin G; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; C023509:ribosomal protein S16; C041184:ribosomal protein S18; C512838:rpsM protein, E coli; D004139:Dinitrofluorobenzene