Ribosomal proteins of Escherichia coli. II. Chemical and physical characterization of the 30 S ribosomal proteins

Craven, Gary R.; Voynow, P.; Hardy, Simon; Kurland, C. G.

doi:10.1021/bi00835a032

Cited by 169 publications

(63 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The apparently selective adsorption of P-Ag by some particles in the cell-free ribosomal preparations also supports the view that the complex was not dissociated during the experiments, and may reflect the different types of ribosome used or the: variation of ribosomal constituents noted by Craven, Voynow, Hardy & Kurland (1969). The fact that some bacterial ribosomes adsorbed P-Ag suggests that aspects of host-parasite specificity involving fungal pigment are not expressed at the ribosomal level.…”

Section: Discussionsupporting

confidence: 63%

The Adsorption of Silver-labelled Fungal Pigment by Bacterial and Apple Plant Ribosomes

Hignett

1973

Journal of General Microbiology

View full text Add to dashboard Cite

S U M M A R YExtracellular material produced by Venturia inaequalis (the apple scab disease organism) formed a tightly bound complex with silver. The silver complex (in aqueous solution) was taken up by whole detached apple leaves (cultivar MM 109) and was observed in the petiole cytoplasm as ribosome-like particles. Diffuse deposits of silver were observed in the cell walls.The silver complex was also taken up by a proportion of the particles present in ribosome preparations made from apple-leaf tissues and from bacterial cultures. Such complexed ribosomes were apparently swollen by internal adsorption of the substance, some showing (in the electron microscope) a faint halo round the central core; the cleft between the sub-units was visible. Comparatively few ribosomes from Pseudomonas rnors-prunorurn showed strong affinity for the silver complex. The ribosomal particles were similar in most respects to the particles seen in treated leaf petioles. Silver-complexed particles prepared from isolated ribosomes were degraded by 'Pronase' to ragged skeletal remnants which were further digested by pancreatic ribonuclease. The latter enzyme did not attack intact particles.The results support evidence from earlier work that the progress of the disease is partly controlled by the effects of fungal pigments on host solute transport (possibly mediated via the cell-wall membrane system) and on hormone-controlled plant metabolism and photosynthesis (via the protein-synthesizing system).

show abstract

Section: Discussionsupporting

confidence: 63%

The Adsorption of Silver-labelled Fungal Pigment by Bacterial and Apple Plant Ribosomes

Hignett

1973

Journal of General Microbiology

View full text Add to dashboard Cite

show abstract

“…Protein S3 had in some but not all runs been eluted from CM-cellulose in two peaks. The proteins in these two peaks moved a t two slightly different positions in two-dimensional electrophoresis and corresponded [I21 to proteins 5 and 9 described by Craven et al [7].…”

mentioning

confidence: 52%

“…They had very similar chemical, physical and immunological properties and were considered as two chemical forms, where one is a derivative of the other [7].…”

mentioning

confidence: 99%

Ribosomal Proteins

Hindennach

Stöffler

Wittmann

1971

European Journal of Biochemistry

181

View full text Add to dashboard Cite

All 21 ribosomal proteins from 3 0 s subunits of Escheriehia coli were isolated in a pure state by the following methods: separation of subunits by zonal centrifugation in B XV rotors, extraction of proteins, CM-cellulose chromatography with pyridine formate gradients, gel filtration on Sephadex G-100 and, in some cases, preparative polyacrylamide block electrophoresis. The purity of isolated proteins was 97-100°/, shown by disc electrophoresis a t pH 4.5 in urea, by two-dimensional polyacrylamide electrophoresis, by dodecylsulfate gel electrophoresis and by cellulose acetate gel electrophoresis. The yield of the isolated proteins was 7 -110 mg depending on the protein and the number of purification steps.Fractionation of total 3 0 s proteins with ammonium sulfate is recommended for quick isolation of proteins S15 and 520 in large quantities.Waller [ 11 showed that starch gel electrophoresis separates the protein moiety of Escherichia eoli ribosomes into many bands. Later studies [2-101 demonstrated that the bands correspond to proteins of different physical and chemical structure. There exists also no immunological cross reaction between the individual 30 S proteins [ 111. There is agreement between various groups on the number, namely 21, and on the nomenclature of the ribosomal proteins from the 3 0 s subunits of E. coli [12].Good and reproducible methods for the isolation of ribosomal proteins on a big scale are the prerequisite not only for their chemical and physical characterization but also for protein-chemical studies on their amino sequences. The present paper describes t h e isolation of all 30 S ribosomal proteins of E. eoli in relatively large quantities (7-110 mg) and high purity (97-100°/,). Haan, Germany). After removal of cell debris and alcoa a t 16000 rev./min for 40 min in a Servall SS-34 rotor, ribosomes were sedimented by overnight centrifugation a t 25000rev.lmin in a Spinco 42 rotor. Ribosomes were resuspended in the buffer described above and purified by three cycles of differential centrifugation, the last with a buffer containing 0.5 M NH,C1 in 0.02 M Tris-HC1 pH 7.8,0.002 M MgC1, and 0.01 M mercaptoethanol. For our first isolations of ribosomes a buffer [14] containing 0.6 M (NH,),SO, instead of 0.5MNH4C1 was used for the last two cycles. MATERIAL AND METHODS IsolationFor separation of subunits aliquots of about 1.5 g ribosomes in 30 ml were dialyzed overnight against 5 1 of 0.01 M potassium phosphate pH 7.4. 0.001 hl MgCl, and 0.01 M mercaptoethanol and centrifuged in Spinco B XV Ti zonal rotors for 17 h a t 25000 rev./ min. A gradient of 7.5O/, to 38O/, sucrose in phosphate buffer and 600 ml overlay was used [15]. Absorbance was recorded a t 295 nm in a flow cell of a Beckman DB spectrophotometer. Subunits were precipitated either with ammonium sulfate (2/3 saturation a t 4 "C)

show abstract

“…This acidic protein cannot be released from the ribosomes by washing the subparticles with solutions of 0.5 M NH4Cl [ 1] or (NH4)$04 [6] . These are conditions used to release supernatant enzymes such as ribonuclease 1 [ 171, or T-and G elongation factors [ 181.…”

Section: Discussionmentioning

confidence: 99%