scholarly journals Modification of human prekeratin during epidermal differentiation

1981 ◽  
Vol 199 (1) ◽  
pp. 145-154 ◽  
Author(s):  
P E Bowden ◽  
W J Cunliffe
Keyword(s):  
High Resolution ◽  
Anatomical Variation ◽  
Sodium Dodecyl ◽  
Complete Removal ◽  
Epidermal Differentiation ◽  
Citrate Buffer ◽  
Molecular Weight Range ◽  
Gradient Gel Electrophoresis ◽  
Size Heterogeneity ◽  
Polypeptide Chains

The polypeptide-chain components of human epidermal prekeratin and keratin were analysed by high-resolution SDS (sodium dodecyl sulphate)/polyacrylamide-gradient-gel electrophoresis. Size heterogeneity existed amongst prekeratin components and at least ten polypeptides, in the molecular-weight range 46,000-70,000, were observed in 0.1 M-citric acid/sodium citrate buffer (pH 2.65) extracts of scale epidermis. Prekeratin from scalp pilosebaceous ducts was identical with that from the contiguous epidermis, and no prekeratin was found in extracts of scale dermis. Prekeratin from plantar epidermis contained additional polypeptide chains, but only slight anatomical variation existed between the non-callus sites examined. Keratin differed from prekeratin in at least two major respects: (a) many major components did not co-electrophorese on high-resolution SDS/polyacrylamide slab gels, and (b) keratin, but not prekeratin, required denaturing and reducing conditions for extraction. Keratin extracted from scale epidermis after complete removal of prekeratin was identical with forearm stratum-corneum keratin. Palmar and plantar keratin contained additional polypeptide chains and had a different size distribution compared with forearm and scalp keratin components. Modification of prekeratin components to produce the keratin polypeptide profile occurred during epidermal differentiation, and these changes appeared to take place in the granular-layer region of the epidermis.

scholarly journals CHEMICAL CHARACTERIZATION OF ISOLATED EPIDERMAL DESMOSOMES

1974 ◽  
Vol 63 (2) ◽  
pp. 524-530 ◽  
Author(s):  
Christine J. Skerrow ◽  
A. Gedeon Matoltsy
Keyword(s):  
Gradient Centrifugation ◽  
Lipid Fraction ◽  
Sodium Dodecyl ◽  
Chemical Characterization ◽  
Citrate Buffer ◽  
Molecular Weight Range ◽  
Sialic Acid Content ◽  
Molecular Weights ◽  
Desmosomal Protein

Desmosomes, isolated from cow nose epidermis by a method utilizing citrate buffer pH 2.6 and density gradient centrifugation, have been analyzed and found to contain approximately 76% protein, 17% carbohydrate, and 10% lipid. Nonpolar amino acids predominate in desmosomal protein, representing 456 residues per 1,000. The sialic acid content is 5 nM/mg of protein. The lipid fraction is composed of approximately 40% cholesterol and 60% phospholipids. Desmosomes are completely solubilized by incubation with 2% sodium dodecyl sulphate and 1% ß-mercaptoethanol. Gel electrophoresis of the denatured desmosomal proteins reveals 24 bands, with mobilities corresponding to a molecular weight range of 15,000–230,000 daltons. Seven of these are considered to be major bands, together constituting 81% of the desmosomal protein. Bands 1 and 2, of molecular weights 230,000 and 210,000 daltons, together comprise 28% by weight of the desmosome. It is suggested that these protein chains are located in the desmosomal plaque. Bands 3 and 4 are PAS-positive, constitute 23% of the desmosomal protein, and have apparent molecular weights of 140,000 and 120,000 daltons, respectively. At least part of this material must originate from the carbohydrate-containing layer which is demonstrated, by histochemistry, to be present in the desmosomal interspace. The possible nature and origin of the remaining major bands, of molecular weights 90,000, 75,000, and 60,000 daltons, are discussed.

scholarly journals Affinity purification and subunit structures of β-N-acetylhexosaminidases A and B from boar epididymis

1984 ◽  
Vol 219 (3) ◽  
pp. 1009-1015 ◽  
Author(s):  
H C Parkes ◽  
J L Stirling ◽  
P Calvo
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Affinity Purification ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Gradient Gel Electrophoresis ◽  
Sepharose 4B ◽  
Electrophoretic Transfer ◽  
Peptide Maps

beta-N-Acetylhexosaminidase from boar epididymis was separated into two forms, A and B, on DEAE-cellulose. Both these forms were excluded from Sepharose S-200 and had apparent Mr values of 510 000 on gradient gel electrophoresis under non-denaturing conditions. Affinity chromatography on 2-acetamido-N-(6-aminohexanoyl)-2-deoxy-beta-D-glucopyranosylam ine coupled to CNBr-activated Sepharose 4B was used to separate and purify beta-N-acetylhexosaminidases A and B that had specific activities of 115 and 380 mumol/min per mg of protein respectively. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of denatured beta-N-acetylhexosaminidase A gave a single major component of Mr 67 000. beta-N-Acetylhexosaminidase B also had this component, and in addition had polypeptides of Mr 29 000 and 26 000. All these polypeptides were glycosylated. Antiserum to the B form precipitated form A from solution and reacted with the 67 000-Mr component or form A after electrophoretic transfer from sodium dodecyl sulphate/polyacrylamide gels to nitrocellulose sheets. The 67 000-Mr components of forms A and B yielded identical peptide maps when digested with Staphylococcus aureus V8 proteinase, and the 29 000-Mr and 26 000-Mr components in form B may be related to the 67 000-Mr polypeptide.

Characterization of Indonesia Decellularized Liver Cubes Scaffold using Scanning Electron Microscopy

2021 ◽  
Vol 52 ◽  
pp. 38-46
Author(s):  
A.A. Ayu Asri Prima Dewi ◽  
Radiana D. Antarianto ◽  
Jeanne Adiwinata Pawitan
Keyword(s):  
Liver Tissue ◽  
Sodium Dodecyl ◽  
Complete Removal ◽  
Sem Analysis ◽  
Ethylene Glycol Tetraacetic Acid ◽  
Native Liver ◽  
Dna Materials ◽  
Protocol Study ◽  
Syringe Injection ◽  
Scanning Electron

Liver biological scaffold was developed in order to resemble native liver tissue environment. It can be achieved by decellularizing native liver tissue that will remove cells and preserve extracellular matrix (ECM). Furthermore, ECM fibers are arranged in a special pattern, which affect liver cell polarity and topography that are important for cells’ implantation, proliferation and differentiation. Therefore, the aim of this study was to evaluate liver cube scaffold topography that was decellularized with fixed multiple sites syringe injection (Indonesia patent number: S00201907930).Rat liver cubes (n=3) underwent decellularization with Ethylene Glycol Tetraacetic Acid (EGTA) immersion and increased Sodium Dodecyl Sulfate (SDS) concentrations using previous multiple sites syringe injection protocol study. Deoxyribonucleic Acid (DNA) concentrations were measured to confirm less DNA materials remaining in scaffolds. Scanning Electron Microscope (SEM) analysis of scaffolds were conducted for topographic characterization compared to undecellularized liver control. Molecular analysis of DNA concentration showed complete removal of DNA material. SEM analysis gave appearance of intact liver cube scaffold microarchitecture. Liver cubes decellularization using multiple sites syringe injection showed good topographic liver scaffold characterization.

Cloning of trg, a gene for a sensory transducer in Escherichia coli

1982 ◽  
Vol 152 (1) ◽  
pp. 372-383
Author(s):  
S Harayama ◽  
P Engström ◽  
H Wolf-Watz ◽  
T Iino ◽  
G L Hazelbauer
Keyword(s):  
Escherichia Coli ◽  
High Resolution ◽  
Sodium Dodecyl Sulfate ◽  
Structural Gene ◽  
Preliminary Analysis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gels ◽  
Physiological Functions ◽  
The Common ◽  
Dependent Modification

Clones of trg, a gene which codes for a chemotactic transducer, were isolated linked to ColE1 and pBR322 vectors. Studies with the hybrid plasmids demonstrated unequivocally that trg is the structural gene for methyl-accepting chemotaxis protein III. The Trg protein was found to be structurally complex, electrophoresing as a series of seven bands on high-resolution sodium dodecyl sulfate-polyacrylamide gels. The multiplicity of bands is a function of the activity of cheR, which codes for a methyltransferase, and of cheB, which codes for a demethylase. It appears that Trg, a quantitatively minor transducer, resembles the two major transducer proteins, Tsr and Tar, in that all three are multiply methylated and also multiply modified in a second way which requires an active cheB gene. However, preliminary analysis of the Trg protein indicated that it is significantly less related structurally to the Tsr or Tar protein than those two transducers are to each other. This implies that the features of multiple methylation and cheB-dependent modification are likely to be critical for the common physiological functions in chemotactic excitation and adaptation performed by all three transducers.

Partial characterization of an abnormal lactate dehydrogenase isoenzyme, LDH-1ex, in serum from a patient with hepatocellular carcinoma.

1989 ◽  
Vol 35 (5) ◽  
pp. 844-848
Author(s):  
D L Kalpaxis ◽  
E E Giannoulaki
Keyword(s):  
Hepatocellular Carcinoma ◽  
Lactate Dehydrogenase ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Chromatography ◽  
Heat Stable ◽  
Single Polypeptide ◽  
Polypeptide Chains

Abstract Serum from a patient with hepatocellular carcinoma contained an abnormal isoenzyme of lactate dehydrogenase (LDH; EC 1.1.1.27), LDH-1ex, that on electrophoresis on 10-g/L agarose gel migrated anodally to the LDH-1 band. This isoenzyme was partly purified by ultrafiltration and preparative electrophoresis. Gel chromatography and sodium dodecyl sulfate/polyacrylamide gel electrophoresis studies of the resulting LDH-1ex preparation suggested that this isoenzyme is probably a tetramer made up of four single polypeptide chains (monomers), each having a molecular mass of about 32,000 Da. LDH-1ex was heat stable and reacted more readily with 2-hydroxybutyrate than did the slower migrating LDH-4 and LDH-5 isoenzymes. LDH-1ex showed no activity when lactate was omitted from the substrate solution or replaced by ethanol.

scholarly journals Interleukin-1 induces interleukin-6 production in peripheral blood monocytes

Blood ◽  
1990 ◽  
Vol 75 (6) ◽  
pp. 1305-1310 ◽  
Author(s):  
G Tosato ◽  
KD Jones
Keyword(s):  
Endothelial Cells ◽  
Interleukin 6 ◽  
Peripheral Blood ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Relative Molecular Mass ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Potent Inducer ◽  
Size Heterogeneity

Abstract Interleukin-6 (IL-6), a multifunctional cytokine produced in monocytes, fibroblasts, endothelial cells, and keratinocytes, is induced by a variety of stimulating signals, including lipopolysaccharide (LPS), poly (I), poly (C), IL-1, tumor necrosis factor (TNF), and platelet- derived growth factor. Some of these signals induce IL-6 effectively only in one cell type, and this selectivity of induction may explain selectivity of biologic effects. In the present study, we show that IL- 1 beta, previously known to be a potent inducer of IL-6 in fibroblasts, endothelial cells, and keratinocytes, but not in monocytes, is also a potent inducer of IL-6 in peripheral blood monocytes. High level IL-6 activity that could be neutralized by specific antibodies to IL-6 was detected in supernatants of IL-1-stimulated monocytes. Maximal induction required IL-1 concentrations of 10 ng/mL. As judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, IL-6 species of relative molecular mass of 19 to 26 Kd could be specifically immunoprecipitated from supernatants of IL-1- induced monocytes. Size heterogeneity is a reported feature of IL-6 produced in a variety of cell types, and monocyte-derived IL-6 induced by either IL-1 or LPS displayed similar size heterogeneity. The highly purified recombinant IL-1 beta preparation used contained little, if any, LPS. In addition, monocyte production of IL-6, induced by IL-1 beta, was specifically neutralized by anti-IL-1 beta antibodies, demonstrating that IL-1, rather than a contaminant in the IL-1 preparation, was responsible for IL-6 induction. A number of biologic activities have been ascribed both to IL-1 and IL-6. The finding that IL-1 induced IL-6 in monocytes may help in defining the spectrum of biologic activities of each of these interactive cytokines.

Ultrathin-layer sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis and silver staining of urinary proteins

1986 ◽  
Vol 7 (11) ◽  
pp. 496-505 ◽  
Author(s):  
Hans-Walter Schiwara ◽  
Thomas Hebell ◽  
Hartmut Kirchherr ◽  
Wilhelm Postel ◽  
Johannes Weser ◽  
...  
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Silver Staining ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Urinary Proteins ◽  
Dodecyl Sulfate ◽  
Gradient Gel Electrophoresis ◽  
Ultrathin Layer

Urease. X. A study by gel electrophoresis of its dissociation by sodium dodecyl sulfate

1969 ◽  
Vol 47 (10) ◽  
pp. 989-991 ◽  
Author(s):  
D. P. Blattler ◽  
George Gorin
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Disulfide Bonds ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Dodecyl Sulfate ◽  
Polypeptide Chains

Urease, m.w. 480 000, treated with an excess of sodium dodecyl sulfate is converted to a product of greatly increased mobility in polyacrylamide gel electrophoresis. We estimate its weight to be about 80 000. Treatment with excess thiol and detergent yielded the same product as detergent alone, indicating that the subunit or subunits do not contain polypeptide chains linked by disulfide bonds.

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