Protein chromatography on adsorbents with hydrophobic and ionic groups. Chromatography of dodecyl sulphate-solubilized proteins of the human erythrocyte membrane on N-(3-carboxypropionyl)aminodecyl-sepharose

Human erythrocyte ‘ghosts’ were solubilized in 0.5% (w/v) sodium dodecyl sulphate at pH 4.0(I = 0.012 mol/I). At a loading of 1-2 mg of protein/ml of column volume, all of membrane proteins were adsorbed to a column of CPAD [N-(3-carboxypropionyl)-aminodecyl]-Sepharose at pH 4.0 (I = 0-012 mol/1) and room temperature (22 degrees C). Many proteins were subsequently desorbed by raising the pH or by including sodium dodecyl sulphate continuously in the eluting buffer. Experiments with a series of adsorbents homologous with CPAD-Sepharose, in which the length of the hydrocarbon chain was varied, provided strong evidence of hydrophobic interactions, in addition to ionic interactions, in the binding of these proteins to CPAD-Sepharose. Elution with increasing-pH gradients at different concentrations of sodium dodecyl sulphate showed that glycophorin (the major sialoglycoprotein) was eluted in the void volume, at recoveries close to 100%, when the detergent concentration was greater than or equal to 0.3% (w/v). Protein E, the major protein, was desorbed late in the pH gradient even at a high (0.5%, w/v) concentration of the detergent, and was always incompletely desorbed, the maximum recovery recorded being 40%. Spectrin (the high-molecular-weight polypeptide pair) did not behave in a well-defined manner, and was found widely distributed among the effluent fractions under all the conditions that were tested.

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Protein chromatography on adsorbents with hydrophobic and ionic groups. Purification of human erythrocyte glycophorin

Biochemical Journal ◽

10.1042/bj1630397 ◽

1977 ◽

Vol 163 (2) ◽

pp. 397-400 ◽

Cited By ~ 12

Author(s):

R J Simmonds ◽

R J Yon

Keyword(s):

Sodium Dodecyl Sulphate ◽

Human Erythrocyte ◽

Sodium Dodecyl ◽

Erythrocyte Ghosts ◽

Electrophoretic Band ◽

Protein Chromatography ◽

Ionic Groups ◽

A Subunit ◽

Triton X

Human erythrocyte glycophorin was purified rapidly by (a) chromatography of a Triton X-100 extract of erythrocyte ‘ghosts’ on N-(3-carboxypropionyl)aminodecyl-Sepharose in buffers containing Triton X-100 or sodium dodecyl sulphate, or (b) chromatography of whole ‘ghosts’, solubilized in sodium dodecyl sulphate, on dodecyl-Sepharose, in buffers containing sodium dodecyl sulphate. The products contained 85-95% glycophorin (electrophoretic band PAS-1) and the major contaminants were glycoproteins PAS-2 (possibly a subunit of glycophorin) and PAS-3.

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The outer membrane of Haemophilus influenzae type b: cell envelope associations of major proteins

Canadian Journal of Microbiology ◽

10.1139/m83-046 ◽

1983 ◽

Vol 29 (2) ◽

pp. 280-287 ◽

Cited By ~ 6

Author(s):

J. W. Coulton ◽

D. T. F. Wan

Keyword(s):

Molecular Weight ◽

Sodium Dodecyl Sulphate ◽

Cell Envelope ◽

Sodium Dodecyl ◽

Haemophilus Influenzae Type ◽

Major Protein ◽

Haemophilus Influenzae Type B ◽

Type B ◽

Molecular Weights ◽

Triton X

Membrane proteins fom the cell envelope of Haemophilus influenzae type b ATCC 9795 were examined by sodium dodecyl sulphate – polyacrylamide gel electrophoresis. When envelopes were extracted with a phosphate-based buffer containing 2% Triton X-100, a major protein of molecular weight 43 000 was detected in fractions containing cytoplasmic membrane proteins. The cell wall material which was Triton X-100 insoluble contained six major proteins of molecular weights 46 000, 40 000, 36 000, 30 000, 27 000, and 16 000. One of these proteins showed a shift in molecular weight from 27 000 to 36 000 when it was heated over a temperature range from 50 °C to 100 °C in buffer containing 2% sodium dodecyl sulphate, 5% 2-mercaptoethanol. This alteration in mobility could be demonstrated either by the membrane-bound form of the protein or by a detergent-soluble form of the protein. Enriched preparations of the 36 000 molecular weight form were obtained by a series of purification steps. Extraction of the Triton X-100 insoluble material with buffer containing 2% Triton X-100, 5.0 mM EDTA yielded chiefly one major protein molecular weight 30 000 and many minor protein species. Pretreatment of the Triton X-100 insoluble fraction with lysozyme followed by extraction with buffer containing 2% Triton X-100, 5.0 mM EDTA released two proteins of molecular weights 16 000 and 27 000 and few minor proteins. By these operational manipulations, the proteins of molecular weights 16 000 and 27 000 may be considered as peptidoglycan-associated proteins.

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Collagen fibril formation in the presence of sodium dodecyl sulphate

Biochemical Journal ◽

10.1042/bj2280551 ◽

1985 ◽

Vol 228 (3) ◽

pp. 551-556 ◽

Cited By ~ 20

Author(s):

G W Dombi ◽

H B Halsall

Keyword(s):

Conformational Changes ◽

Sodium Dodecyl Sulphate ◽

Collagen Fibril ◽

Hydrophobic Interactions ◽

Sodium Dodecyl ◽

Distal Region ◽

Fibril Formation ◽

Collagen Fibrillogenesis ◽

Fibril Morphology

Sodium dodecyl sulphate (SDS) was used to weaken both the electrostatic and the hydrophobic interactions during collagen fibrillogenesis in vitro. The rate and extent of fibril formation as well as fibril morphology were affected by SDS concentration. Both the formation of large fibrils at 0.3 mM-SDS and the complete cessation of fibril formation at 0.5 mM-SDS were considered to be the result of SDS-induced conformational changes in the non-helical telopeptides. A possible mechanism of SDS interaction with the N-terminal and the distal region of the C-terminal telopeptides is offered.

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Some observations on the choice of detergent for solubilization of the human erythrocyte membrane

Biochemical Journal ◽

10.1042/bj1640465 ◽

1977 ◽

Vol 164 (2) ◽

pp. 465-468 ◽

Cited By ~ 28

Author(s):

D A W Grant ◽

S Hjertén

Keyword(s):

Erythrocyte Membrane ◽

Sodium Dodecyl Sulphate ◽

Human Erythrocyte ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Human Erythrocyte Membrane ◽

Ionic Detergents ◽

Triton X

Solubilization of the human erythrocyte membrane by seven detergents is described. Components released into the supernatant or retained in the residue were identified by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Two non-ionic detergents exhibiting little u.v. absorption were more efficient than u.v.-absorbing Triton X-100. Evidence is presented of an interchange between protein PAS 1 and protein PAS 2.

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Chick embryonic pigmented retina is one of the group of epithelioid tissues that lack cytokeratins and desmosomes and have intermediate filaments composed of vimentin

Journal of Cell Science ◽

10.1242/jcs.71.1.61 ◽

1984 ◽

Vol 71 (1) ◽

pp. 61-74

Author(s):

R.J. Docherty ◽

J.G. Edwards ◽

D.R. Garrod ◽

D.L. Mattey

Keyword(s):

Retinal Pigment Epithelium ◽

Intermediate Filaments ◽

Vascular Endothelium ◽

Sodium Dodecyl Sulphate ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Sodium Dodecyl ◽

Major Protein ◽

Ultrastructural Studies ◽

Chick Embryonic Tissue

Using sodium dodecyl sulphate/polyacrylamide gels to analyse detergent-insoluble residues, and indirect immunofluorescence, we have found that the major protein of intermediate filaments in cultures and freshly explanted fragments of chick embryonic retinal pigment epithelium (RPE) is vimentin. Moreover, these cells also fail to stain with antibodies against cytokeratins and most components of true desmosomes (maculae adhaerentes). Staining with anti-vinculin antibody suggests that the principal intercellular junction is the zonula adherens. Thus although RPE is an epithelium according to all other criteria, it belongs to a group of tissues (including vascular endothelium, iris and lens-forming epithelium) that have intermediate filaments composed of vimentin and possess neither cytokeratins nor desmosomes. That a tissue can be fully epithelial by other criteria, whilst lacking these components, is in agreement with other work, which has shown a lack of effect of micro-injection of antibodies to cytokeratin, and of the suppression of desmosome formation, on epithelial organization in culture. Although our observations were made solely on chick embryonic tissue, we suggest that published ultrastructural studies are consistent with the possibility that RPE of other species, including human, may lack true desmosomes.

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Isolation and protein composition of gap junctions from rabbit hearts

Biochemical Journal ◽

10.1042/bj2050189 ◽

1982 ◽

Vol 205 (1) ◽

pp. 189-194 ◽

Cited By ~ 31

Author(s):

C K Manjunath ◽

G E Goings ◽

E Page

Keyword(s):

Gap Junctions ◽

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Protein Composition ◽

Polyacrylamide Gel ◽

Major Protein ◽

Electron Microscopic ◽

Gap Junctional

We have modified a method for isolating gap-junctional membrane from mouse hearts [Kensler & Goodenough (1980) J. Cell Biol. 86, 755-764] to isolate gap junctions of comparable purity from rabbit hearts more rapidly, with better yield, and without resort to non-ionic detergents. Purification was monitored by electron microscopy of thin-sectioned membrane pellets and by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Gap junctions were obtained as vesicles whose mean surface area approximated that of junctions in intact myocardial cells. About 10-20% of the vesicles were ferritin-impermeable. Approx. 125 micrograms of membrane protein was obtained per 8 g of rabbit heart. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of purified gap junctions showed five major protein bands of mol.wts. 46 000, 44 000, 33 000, 30 000 and 28 500 that co-purified with the junctions. This protein composition was nearly identical with that published for gap junctions of mouse hearts, and differed markedly from the protein composition of gap junctions from non-excitable cells (lens and liver). The constancy of junctional protein composition between hearts of two different species and its non-identity with that from liver and lens suggest that, although gap-junctional structure in mammalian tissues seems to be remarkably similar by electron-microscopic techniques, junctional-channel protein composition actually varies from tissue to tissue and may be adapted to the permeability requirements of the tissue.

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Interaction of the myelin basic protein with the anionic detergent sodium dodecyl sulphate

Biochemical Journal ◽

10.1042/bj1690281 ◽

1978 ◽

Vol 169 (2) ◽

pp. 281-285 ◽

Cited By ~ 17

Author(s):

A J S Jones ◽

M G Rumsby

Keyword(s):

Myelin Basic Protein ◽

Sodium Dodecyl Sulphate ◽

Basic Protein ◽

Hydrophobic Interactions ◽

Rapid Development ◽

Sodium Dodecyl ◽

Molecular Size ◽

Transient Increase ◽

Tryptophan Residue ◽

Molar Ratios

The interaction of the myelin basic protein and two peptides derived from it with the anionic detergent SDS (sodium dodecyl sulphate) was studied. At molar ratios of detergent/protein of up to approx. 20:1 the transient increase in turbidity (as measured by increases in A230) is proportional to the ratio. Between ratios of 30:1 and 100:1 the effect of the detergent is constant and maximal. At molar ratios exceeding 100:1 the transient increase in turbidity decreases with increasing amounts of detergent. With increasing ionic strength the rapid development of turbidity is inhibited, whereas the slow decay of turbidity is not affected. Neither of the peptide fragments produced by cleavage of the myelin basic protein at the single tryptophan residue, nor both when mixed, produce measurable turbidity when mixed with SDS. Under similar conditions poly-L-lysine of similar molecular size to the basic protein shows the increase in turbidity but not the decay. The interaction between the protein and SDS is interpreted in molecular terms, which involve the initial ionic interaction of the detergent with protein resulting in aggregation and turbidity in the solution. Within the aggregated complexes molecules rearrange to maximize hydrophobic interactions.

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Electron donor–acceptor complexes between naphthylamines and methyl viologen in aqueous sodium dodecyl sulphate solution

Canadian Journal of Chemistry ◽

10.1139/v86-139 ◽

1986 ◽

Vol 64 (5) ◽

pp. 845-848 ◽

Cited By ~ 9

Author(s):

S. G. Bertolotti ◽

J. J. Cosa ◽

H. E. Gsponer ◽

M. Hamity ◽

C. M. Previtali

Keyword(s):

Electron Donor ◽

Sodium Dodecyl Sulphate ◽

Methyl Viologen ◽

Sodium Dodecyl ◽

Local Concentration ◽

Detergent Concentration ◽

Aqueous Sodium ◽

Sds Micelles ◽

Donor Acceptor ◽

Experimental Values

The electron donor–acceptor (EDA) interaction between methyl viologen (MV2+) and 1-naphthylamine (1NA), 2-naphthylamine (2NA), and N,N-dimethyl-1-naphthylamine (DMA) was studied in water and in aqueous sodium dodecyl sulphate (SDS). The experimental values of the association constants in water were 8.9, 9.8, and 2.8 M−1 for 1NA, 2NA, and DMA, respectively. In the presence of SDS the observed values were very much higher and strongly dependent upon the detergent concentration. The enhancement in the interaction is due to an increase in the local concentration of the partners in the micellar pseudophase. MV2+ itself was shown to strongly interact with the micelles. A new absorption is present at the red tail of the spectrum of MV2+ in the presence of SDS micelles. An association constant of 1700 M−1 was obtained from this absorption.

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Identifikasi Daging Tikus Pada Produk Baso Dengan Metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE)

YARSI Medical Journal ◽

10.33476/jky.v26i2.394 ◽

2018 ◽

Vol 26 (2) ◽

pp. 058

Author(s):

Anna P. Roswiem ◽

Triayu Septiani

Keyword(s):

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Sds Page

Bahan baku untuk membuat baso adalah daging hewan, pada umumnya dari daging sapi, ayam, ikan dan babi. Di beberapa daerah di Indonesia terjadi kasus baso tikus. Tujuan penelitian ini adalah menguji ada tidaknya kandungan daging tikus pada produk baso yang dijual di pasar Cempaka Putih-Kecamatan Kramat Jakarta Pusat dan di pedagang baso atau mie baso di sekitar kampus Universitas YARSI Jakarta. Daging adalah protein salah satu metode untuk mengidentifikasi protein adalah metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Hasil penelitian menunjukkan bahwa dari 6 sampel baso terindikasi ada 2 sampel baso dengan nomor 1 dan 5 yang dibuat dari campuran daging sapi dan tikus; ada 1 sampel baso dengan nomor 6 yang terbuat dari daging tikus; dan 2 sampel baso dengan nomor 2 dan 3 yang terbuat dari campuran sapi dan babi, dan hanya 1 sampel baso dengan nomor sampel 4 yang benar-benar terbuat dari daging sapi.

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