Ribosomal proteins

Kaltschmidt, E.; Stöffler, Georg; Dzionara, M.; Hg, Wittmann

doi:10.1007/bf00267700

Cited by 42 publications

(12 citation statements)

References 14 publications

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“…This exchange could not be detected immunologically, whereas the difference between S7 K and S7 B could be detected (19). Under the experimental conditions used here, both antisera showed complete identity and gave no spur formation.…”

Section: Structural Differences Of the 30s Proteinsmentioning

confidence: 60%

“…Furthermore, it established the different structures of proteins S9 and Si 1, and S16 and S17 (31), two groups of proteins that were difficult to separate. This method further revealed some sequence differences between protein S7 from E. coli strains B and K, but failed to detect the one aminoacid difference between protein S5 from the two strains (19).…”

Section: Discussionmentioning

confidence: 97%

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Sequence Differences of Escherichia coli 30S Ribosomal Proteins as Determined by Immunochemical Methods

Stöffler

Wittmann

1971

Proc. Natl. Acad. Sci. U.S.A.

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112

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Antisera specific for each of the 21 homogenous ribosomal proteins from 30S subunits of Escherichia coli were used to investigate, by immunodiffusion and immunoprecipitation, whether there are any extensive sequence homologies among these proteins.No immunological crossreactions were detected between any of the proteins. Therefore, we conclude that no significant common sequences exist among any of the 21 30S ribosomal proteins of E. coli.The exact features and structural requirements of antigenic determinants have been well characterized for synthetic antigens (1). Since the early studies of Landsteiner (2) on the specificity of serological reactions with natural protein antigens, progress has been made towards elucidating the properties of antigenic determinants in naturally occurring proteins (3, 4). The presence of many specific determinants on a single natural protein induces a heterogenous population of different antibodies, each specific for a different determinant distributed over the antigen protein (reviewed in ref. 5).Our purpose was to isolate antisera specific for each of the 30S proteins of Escherichia coli, so that we could investigate the extent of the structural differences between each protein.We considered this immunological method to be uniquely sensitive to structural details, and only surpassed by determination of the complete aminoacid sequence of each protein. The strategy of the experiments was to establish whether any immunological crossreaction occurred heterologously between individual homogenous 30S subunit proteins and their specific antisera. MATERIALS AND METHODSStrains. E. coli strain K12 (A19) (6) was used unless otherwise stated. E. coli B was used for the preparation of proteins S5 B and S7 B.Ribosomes and Ribosomal Proteins. Cells were grown in rich medium and harvested 2/3 of the way through the log phase. Ribosomes were obtained by differential centrifugation and washed twice with 0.5 M NH4Cl (7) or in 0.6 M (NH4)2SO4 (8). Subunits were separated and single 30S ribosomal proteins were isolated as described (9).The fractionated proteins were identified by 2-dimensional polyacrylamide gel electrophoresis (10, 11). 30-100 ,ug of each protein, together with 300-400 ,ug of total 70S ribosomal proteins, were electrophoresed; and the position of the main spot, relative to the faint background, identified the protein.Total 70S ribosomal protein used for immunochemical methods was prepared either by extraction with 67% acetic acid at 35 mM MgCl2 (8) or by incubating ribosomes with an equal volume of 8 M urea-4 M LiCl for 48 hr at 0C. The supernatant after a 30-min centrifugation was used. No differences were found between the two extraction methods, except that the proteins prepared with LiCl-urea were more soluble in buffers without urea. When total 30S ribosomal protein was used for serological work, the 30S particles were purified by two succeeding zonal centrifugations.Purity and Characterization of Proteins. The following methods were used to estimate the purity of each...

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Section: Structural Differences Of the 30s Proteinsmentioning

confidence: 60%

Section: Discussionmentioning

confidence: 97%

Sequence Differences of Escherichia coli 30S Ribosomal Proteins as Determined by Immunochemical Methods

Stöffler

Wittmann

1971

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

112

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“…TB, the tryptone medium used, was described (16 (24). Proteins were extracted from purified ribosomes by adding.2 vol ofglacial acetic acid (25). The proteins were then precipitated with acetone and washed (26), vacuum dried, and frozen at -700C.…”

Section: Methodsmentioning

confidence: 99%

Evidence that ribosomal protein S10 participates in control of transcription termination.

Friedman

Schauer

Baumann

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

110

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“…The other of the 21 proteins have their isoelectric points at pH > 8.3 ( fig.2), and proteins S 5 and S 7 take positions on the electropherogram characteristic of the proteins separated from strain B of E. coil [5].…”

Section: S Subunit Of Ribosomes From E Coli 113-3mentioning

confidence: 99%

“…Employing this method, Kaltschmidt et al [5] performed an analysis of the ribosomal proteins from E. coli strains B, C, K12 and MRE 600 and showed different physico-chemical properties of proteins $5 and $7 derived from those four strains. Up till now, no map has been made of the ribosomal proteins derived from the E. coli 113-3 mutant, of interest because of their functions in vitamin BI2 synthesis.…”

Section: Introductionmentioning

confidence: 99%

Comparative studies on ribosomal proteins of Escherichia coli 113‐3 and strains B, C and K₁₂

Pezacka-Fojudzka¹,

Walerych²

1976

FEBS Letters

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Ribosomal proteins

Cited by 42 publications

References 14 publications

Sequence Differences of Escherichia coli 30S Ribosomal Proteins as Determined by Immunochemical Methods

Sequence Differences of Escherichia coli 30S Ribosomal Proteins as Determined by Immunochemical Methods

Evidence that ribosomal protein S10 participates in control of transcription termination.

Comparative studies on ribosomal proteins of Escherichia coli 113‐3 and strains B, C and K₁₂

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Ribosomal proteins

Cited by 42 publications

References 14 publications

Sequence Differences of Escherichia coli 30S Ribosomal Proteins as Determined by Immunochemical Methods

Sequence Differences of Escherichia coli 30S Ribosomal Proteins as Determined by Immunochemical Methods

Evidence that ribosomal protein S10 participates in control of transcription termination.

Comparative studies on ribosomal proteins of Escherichia coli 113‐3 and strains B, C and K12

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Comparative studies on ribosomal proteins of Escherichia coli 113‐3 and strains B, C and K₁₂