scholarly journals Precise determination of RNA–protein contact sites in the 50 S ribosomal subunit of Escherichia coli

1998 ◽  
Vol 334 (1) ◽  
pp. 39-42 ◽  
Author(s):  
Bernd THIEDE ◽  
Henning URLAUB ◽  
Helga NEUBAUER ◽  
Gerlinde GRELLE ◽  
Brigitte WITTMANN-LIEBOLD
Keyword(s):  
Escherichia Coli ◽  
Ribosomal Proteins ◽  
Ribosomal Subunit ◽  
Laser Desorption ◽  
Precise Determination ◽  
Cross Linking ◽  
Laser Desorption Ionization ◽  
Precise Data ◽  
Contact Sites

RNA–protein cross-linked complexes were isolated and purified to obtain precise data about RNA–protein contact sites in the 50 S ribosomal subunit of Escherichia coli. N-terminal microsequencing and matrix-assisted laser desorption ionization MS were used to identify the cross-linking sites at the amino acid and nucleotide levels. In this manner the following contact sites of five ribosomal proteins with the 23 S rRNA were established: Lys-67 of L2 to U-1963, Tyr-35 of L4 to U-615, Lys-97 of L21 to U-546, Lys-49 of L23 to U-139 or C-140 and Lys-71 and Lys-74 of L27 to U-2334.

Structural organization of escherichia coli ribosomes as determined by immuno electron microscopy

1978 ◽  
Vol 36 (2) ◽  
pp. 166-167 ◽  
Author(s):  
G. Stöffler ◽  
R.W. Bald ◽  
J. Dieckhoff ◽  
H. Eckhard ◽  
R. Lührmann ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Escherichia Coli ◽  
Ribosomal Proteins ◽  
Structural Organization ◽  
Three Dimensional ◽  
Ribosomal Subunit ◽  
Spatial Arrangement ◽  
Small Subunit ◽  
Antibody Binding ◽  
Immuno Electron Microscopy

A central step towards an understanding of the structure and function of the Escherichia coli ribosome, a large multicomponent assembly, is the elucidation of the spatial arrangement of its 54 proteins and its three rRNA molecules. The structural organization of ribosomal components has been investigated by a number of experimental approaches. Specific antibodies directed against each of the 54 ribosomal proteins of Escherichia coli have been performed to examine antibody-subunit complexes by electron microscopy. The position of the bound antibody, specific for a particular protein, can be determined; it indicates the location of the corresponding protein on the ribosomal surface.The three-dimensional distribution of each of the 21 small subunit proteins on the ribosomal surface has been determined by immuno electron microscopy: the 21 proteins have been found exposed with altogether 43 antibody binding sites. Each one of 12 proteins showed antibody binding at remote positions on the subunit surface, indicating highly extended conformations of the proteins concerned within the 30S ribosomal subunit; the remaining proteins are, however, not necessarily globular in shape (Fig. 1).

scholarly journals Determination of 1,5‒anhydro‒glucitol–carrier protein conjugates by matrix‒assisted laser desorption/ionization tof mass spectrometry and antibody formation

2000 ◽  
Vol 14 (3) ◽  
pp. 85-92 ◽  
Author(s):  
Li-Jiang Xuan ◽  
Hiroyuki Tanaka ◽  
Satoshi Morimoto ◽  
Yukihiro Shoyama ◽  
Hiroshi Akanuma ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Serum Albumin ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Protein Conjugates ◽  
Desorption Ionization ◽  
Chicken Lysozyme ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

In order to prepare the monoclonal antibody against 1,5-anhydro-glucitol (1), it was conjugated with bovine serum albumin (BSA), human serum albumin (HSA) or chicken lysozyme (CL) using succinate orβ-alanine succinate as a spacer to produce individual antigen conjugates. The number of hapten contained in each antigen conjugate was determined by matrix-assisted laser desorption/ionization tof mass (MALDI-tof-MS) spectrometry. The formation of monoclonal antibody (MAb) was also discussed.

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