Membrane Protein Synthesis in Micrococcus lysodeikticus and Selective Effect of Chloramphenicol

1975 ◽  
Vol 53 (5) ◽  
pp. 615-622
Author(s):  
C. Garcia Mendoza ◽  
M. Novaes Ledieu
Keyword(s):  
Protein Synthesis ◽  
Membrane Proteins ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Specific Radioactivity ◽  
Inhibitory Effect ◽  
Selective Effect ◽  
Cytoplasmic Membranes ◽  
Micrococcus Lysodeikticus

Micrococcus lysodeikticus cytoplasmic membranes labeled with [14C]arginine plus [14C]-threonine were prepared and subjected to mild washing treatments to fractionate membrane proteins. Polyacrylamide gel electrophoresis of total membranes, in the presence of sodium dodecyl sulfate, results in the separation of 28–30 bands of labeled protein. Three peaks of protein show higher specific radioactivity than the others. Chloramphenicol at 100 μg/ml inhibits the incorporation of labeled precursors into membrane proteins by 45–70%, some of them being more affected by the antibiotic. From all available results, we suggest that the partial inhibitory effect shown by this antibiotic could be due to the existence of specific biosynthetic sites for some membrane proteins, which are differently affected by chloramphenicol.

A novel Bicine running buffer system for doubled sodium dodecyl sulfate – polyacrylamide gel electrophoresis of membrane proteins

2006 ◽  
Vol 27 (14) ◽  
pp. 2984-2995 ◽  
Author(s):  
Taufika Islam Williams ◽  
Jennifer C. Combs ◽  
Anup P. Thakur ◽  
Herbert J. Strobel ◽  
Bert C. Lynn
Keyword(s):  
Membrane Proteins ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Buffer System ◽  
Dodecyl Sulfate

Outer membrane proteins from Serratia marcescens

1993 ◽  
Vol 39 (1) ◽  
pp. 108-111 ◽  
Author(s):  
Marta Puig ◽  
Carme Fusté ◽  
Miquel Viñas
Keyword(s):  
Membrane Proteins ◽  
Serratia Marcescens ◽  
Outer Membrane ◽  
Nutrient Availability ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Outer Membrane Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cultural Conditions

The outer membrane proteins (OMPs) of several strains of Serratia marcescens have been studied by sodium dodecyl sulphate – urea – polyacrylamide gel electrophoresis. Four major OMPs, named Omp1, Omp2, Omp3, and OmpA (42, 40, 39, and 37 kDa, respectively), have been visualized. The relative proportions of Omp2 and Omp3 depend on cultural conditions (temperature of incubation, osmolarity, and nutrient availability).Key words: Serratia marcescens, outer membrane proteins, porin.

Compositional and electrophoretic changes in buffalo milk-fat globule membrane proteins during lactation

1976 ◽  
Vol 43 (3) ◽  
pp. 381-388 ◽  
Author(s):  
Sukhminder Singh ◽  
N. C. Ganguli
Keyword(s):  
Membrane Proteins ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Milk Fat ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Milk Fat Globule Membrane ◽  
Milk Fat Globule ◽  
Fat Globule

SummaryChemical analyses, polyacrylamide-gel electrophoresis and isoelectric focusing of milk-fat globule membrane proteins (FGMP) obtained from the milk of 2 Murrah buffaloes were done to determine if any change in composition occurred during lactation. Changes in the levels of sialic acid, hexose, hexosamine, N and P were found in the FGMP obtained at different stages of lactation. On the day of parturition, 8 major proteins in FGMP were determined by sodium dodecyl sulphate polyacrylamide-gel electrophoresis whereas 6 major proteins were obtained in FGMP of middle and late lactation milks. Isoelectric focusing of FGMP showed 8–9, 9–13 and 13–16 proteins from colostrum, middle and late lactation milks, respectively and the isoelectric pH of the proteins varied from 5·25 to 7·80, 5·85 to 8·30 and 5·75 to 8·61 respectively.

scholarly journals Synthesis and turnover of plasma-membrane proteins and glycoproteins in a neuroblastoma cell line

1976 ◽  
Vol 154 (1) ◽  
pp. 57-64 ◽  
Author(s):  
R A Mathews ◽  
T C Johnson ◽  
J E Hudson
Keyword(s):  
Steady State ◽  
Membrane Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Surface Membrane ◽  
Neuroblastoma Cell ◽  
Sodium Dodecyl ◽  
Neuroblastoma Cells ◽  
Specific Radioactivity ◽  
Neuroblastoma Cell Line ◽  
Total Membrane

A kinetic analysis of the appearance of 14C-labelled proteins in the surface membranes isolated from exponentially growing neuroblastoma cells (N2a) showed that the total membrane proteins reached a steady-state specific radioactivity in 18-20 h. However, examination of individual protein bands resolved by sodium dodecyl sulphate-urea-polyacrylamide-gel electrophoresis illustrated that differences in the kinetics of specific surface-membrane proteins could be detected. Although most of the protein bands reached a steady-state specific radioactivity at a time similar to that for total membrane proteins, at least two bands (mol. wt. 180000 and 130000) attained the steady-state within 8-10 h. It was shown by the use of dual-labelling techniques that these two protein bands turned over in the surface membranes of neuroblastoma N2a cells at least 180 and 150% faster than the total membrane protein. These two proteins were glycosylated and located on the outer surface of the cells, since they were labelled with radioactive carbohydrates and readily removed by treatment of the intact neuroblastoma cell with proteinases.

Stability of Secretory Granules Containing Growth Hormone and Prolactin from Bovine Anterior Pituitary Gland

1974 ◽  
Vol 52 (4) ◽  
pp. 327-335 ◽  
Author(s):  
André Lemay ◽  
Fernand Labrie ◽  
Denis Drouin
Keyword(s):  
Growth Hormone ◽  
Gel Electrophoresis ◽  
Secretory Granules ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Inhibitory Effect ◽  
High Yield ◽  
Maximal Effect ◽  
Enzymatic Markers ◽  
The Stability

Bovine adenohypophyseal secretory granules were purified by a technique giving a high yield of large granules containing 70–90% of prolactin (PRL) and 10–20% of growth hormone (GH). Purity of the preparation was checked by electron microscopy and enzymatic markers. Stability of the isolated secretory granules was studied by measurement of the optical density of the granule suspensions and by measurement of GH and PRL release by disc gel electrophoresis at pH 8.9. At 0 °C, granules suspended in an hypotonic medium are stable at pH 7.4. At room temperature, they are particularly stable at acidic pH but are almost completely solubilized at a pH ranging from 7.0 to 10.0, a half-maximal effect on PRL release being observed at pH 7.5. Lysolecithin has a marked solubilizing effect at a pH of 3.0–6.0 whereas 0.15% sodium dodecyl sulfate does not affect granule stability at a pH below 5. ATP (0.5 mM) inhibits GH and PRL release to, respectively, 65 and 27% of the control rates during a 60 min incubation at 37 °C and pH 7.4. A concentration of 2 mM Ca2+ has a marked inhibitory effect on both GH and PRL release whereas 2 mM Mg2+ inhibits PRL release but does not affect the release of GH. EGTA increases PRL release by 30% but does not significantly affect GH release. As evidenced by polyacrylamide gel electrophoresis, no degradation of GH or PRL occurred during a 1 h incubation of secretory granules at 37 °C and pH 7.4. These data suggest a role of Ca2+- and Mg2+-ATPase activities on the stability and possibly the formation of GH- and PRL-containing secretory granules.

Products of mitochondrial protein synthesis in Tetrahymena

1979 ◽  
Vol 57 (4) ◽  
pp. 314-320 ◽  
Author(s):  
Paul G. Young ◽  
Neil P. Hunter
Keyword(s):  
Protein Synthesis ◽  
Gel Electrophoresis ◽  
Trichloroacetic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Mitochondrial Protein ◽  
Sodium Dodecyl ◽  
Apparent Molecular Weight ◽  
Mitochondrial Proteins ◽  
Mitochondrial Protein Synthesis ◽  
Chloroform Methanol

The products of mitochondrial protein synthesis have been investigated in Tetrahymena after labelling with [35S]methionine in the presence of cycloheximide. The labelled proteins were analyzed by sodium dodecyl sulfate slab polyacrylamide gel electrophoresis. We have identified 13 electrophoretically discrete bands as well as 4 other bands with a more variable occurrence. These proteins ranged in apparent molecular weight from 8100 to 57 500. The cycloheximide-resistant incorporation could be blocked with chloramphenicol. The mitochondrial proteins appeared to be in a disaggregated state and were stable to agents such as trichloroacetic acid (hot or cold) and chloroform–methanol. The pattern of proteins was similar following labelling times ranging from 30 min to 3 h.

scholarly journals Purification and properties of different forms of modeccin, the toxin of Adenia digitata. Separation of subunits with inhibitory and lectin activity

1980 ◽  
Vol 185 (1) ◽  
pp. 203-210 ◽  
Author(s):  
L Barbieri ◽  
M Zamboni ◽  
L Montanaro ◽  
S Sperti ◽  
F Stirpe
Keyword(s):  
Protein Synthesis ◽  
Affinity Chromatography ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Purification And Properties ◽  
Inhibitor Of Protein Synthesis ◽  
Sepharose 4B

1. The subunits were isolated of modeccin (subsequently referred to as modeccin 4B), the toxin purified from the roots of Adenia digitata by affinity chromatography on Sepharose 4B [Gasperi-Campani, Barbieri, Lorenzoni, Montanaro, Sperti, Bonetti & Stirpe (1978) Biochem J. 174, 491-496]. They are an A subunit (mol.wt. 26 000), which inhibits protein synthesis, and a B subunit (mol.wt. 31 000), which binds to cells. Both sununits, as well as intact modeccin, gave single bands on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, but showed some heterogeneity on isoelectric focusing and on polyacrylamide-gel electrophoresis at pH 9.5. 2. A second form of modeccin, not retained by Sepharose 4B, was purified by affinity chromatography on acid-treated Sepharose 6B: this form is subsequently termed modeccin 6B 3. Modeccin 6B has a molecular weight indistinguishable from that of modeccin 4B, and consists of two subunits of mol.wts. 27 000 and 31 000, joined by a disulphide bond. The subunits were not isolated because of their high insolubility in the absence of sodium dodecyl sulphate. 4. As compared with modeccin 4B, modeccin 6B is slightly less toxic to animals, does not agglutinate erythrocytes, and is a more potent inhibitor of protein synthesis in a lysate of rabbit reticulocytes, giving 50% inhibition at the concentration of 0.31 microgram/ml.

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