Precise determination of RNA-protein contact sites in the 50 S ribosomal subunit of Escherichia coli.
نویسندگان
چکیده
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.
منابع مشابه
Immunogenicity of a Fusion Protein Comprising Coli Surface Antigen 3 and Labile B Subunit of Enterotoxigenic Escherichia coli
Background: Enterotoxigenic Escherichia coli (ETEC) strains are the major causes of diarrheal disease in humans and animals. Colonization factors and enterotoxins are the major virulence factors in ETEC pathogenesis. For the broad-spectrum protection against ETEC, one could focus on colonization factors and non-toxic heat labile as a vaccine candidate. Methods: A fusion protein is composed of a...
متن کاملIncorporation of dinitrophenyl protein L23 into totally reconstituted Escherichia coli 50 S ribosomal subunits and its localization at two sites by immune electron microscopy.
Escherichia coli ribosomal protein L23 was derivatized with [3H]2, 4-dinitrofluorobenzene both at the N terminus and at internal lysines. Dinitrophenyl-L23 (DNP-L23) was taken up into 50 S subunits from a reconstitution mixture containing rRNA and total 50 S protein depleted in L23. Unmodified L23 competed with DNP-L23 for uptake, indicating that each protein form bound in an identical or simil...
متن کاملA ribonucleoprotein core in the 50 S ribosomal subunit of Escherichia coli.
Many methods have been used for investigating the structural organisation of the ribosome. Although steady progress has been made in characterising both the RNA and protein regions that are accessible on the surface of the ribosome and the approximate locations of the RNA binding sites of the proteins, little is known about the internal organisation of proteins and RNA in the ribosomal subunits...
متن کاملConstruction and Expression of a Fused Gene for B Subunit of the Heat-Labile and a Truncated Form of the Heat-Stable Enterotoxins in Escherichia coli
Elaboration of different toxins by enterotoxigenic E. coli has been considered as one of the main virulence factors contributing to the manifestation of disease caused by these microorganisms. Various strategies have been employed to raise antibodies against these toxins as a line of defense. In this study, the 3’ terminus of the gene that codes for the binding subunit of the heat-labile entero...
متن کاملExpression of Recombinant Protein B Subunit Pili from Vibrio Cholera
Background & Aims: Vibrio cholerae is a gram-negative bacterial pathogen that causes cholera disease. Following ingestion by a host and entry into the upper intestine, V. cholera colonizes and begins to emit enterotoxin. One of the most pathogenic factors of Vibrio cholera is toxin-coregulated pili (TCP). ToxinCoregulated pili is as the primary factor requiered for the colonization and insisten...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- The Biochemical journal
دوره 334 ( Pt 1) شماره
صفحات -
تاریخ انتشار 1998