scholarly journals Purification and properties of plaice metallothionein, a cadmium-binding protein from the liver of the plaice (Pleuronectes platessa)

1979 ◽  
Vol 183 (2) ◽  
pp. 277-283 ◽  
Author(s):  
J Overnell ◽  
T L Coombs
Keyword(s):  
Positive Reaction ◽  
Binding Protein ◽  
Periodic Acid ◽  
Molar Ratio ◽  
Amino Acid Residues ◽  
Thiol Groups ◽  
Single Chain ◽  
Periodic Acid Schiff ◽  
Pleuronectes Platessa ◽  
Mammalian Metallothioneins

A low-molecular-weight protein induced in the liver of the plaice (Pleuronectes platessa) by exposure to cadmium was purified and characterized. It is closely similar to mammalian metallothioneins in all of its properties in that it is a single-chain cadmium-binding protein of approx. 7000 mol.wt. with a high cysteine content (31 mol%) and no aromatic amino acid residues. The thiol groups of the cysteine residues complex with the cadmium in a SH/Cd molar ratio of 3:1 and produce a characteristic absorption maximum at 250 nm. Unlike the mammalian metallothioneins, however, metal analyses reveal only traces of zinc and copper in addition to cadmium. The presence of carbohydrate previously assumed from a positive reaction with periodic acid/Schiff reagent has now been disproved, and the positive reaction attributed to interaction with the thiol groups in the protein.

scholarly journals On the association of glycoprotein Ib and actin-binding protein in human platelets.

1985 ◽  
Vol 100 (1) ◽  
pp. 317-321 ◽  
Author(s):  
J R Okita ◽  
D Pidard ◽  
P J Newman ◽  
R R Montgomery ◽  
T J Kunicki
Keyword(s):  
Binding Protein ◽  
Periodic Acid ◽  
Human Platelets ◽  
Actin Binding ◽  
Affinity Column ◽  
Periodic Acid Schiff ◽  
Actin Binding Protein ◽  
Coomassie Blue ◽  
Sds Page ◽  
Immunoblot Technique

Glycoprotein (GP) Ib was purified from lysates of human platelets prepared in the presence or absence of inhibitors of the endogenous calcium-activated neutral protease (CANP) by immunoaffinity chromatography, employing the GPIb-specific murine monoclonal antibody, AP1, coupled to Sepharose CL4B. When derived from lysates prepared in the presence of EDTA or leupeptin, the eluate from the AP1-affinity column contained a 240,000-260,000-mol-wt protein in addition to GPIb. In SDS PAGE, this protein was stained by Coomassie Blue R, but not by the periodic acid-Schiff reagent, and it was not labeled with 125I in intact platelets by the lactoperoxidase-catalyzed method. When derived from lysates prepared in the absence of CANP inhibitors, the eluate contained only GPIb and its proteolytic derivative, glycocalicin. A change in the electrophoretic mobility of GPIb consistent with its association with the 240,000-260,000-mol-wt protein was confirmed by crossed immunoelectrophoresis. By an immunoblot technique involving transfer of proteins eluted from the AP1-affinity column and separated by SDS PAGE onto a nitrocellulose membrane, the 240,000-260,000-mol-wt protein bound polyclonal goat antibody raised against rabbit macrophage actin-binding protein (ABP). On the basis of these results, we conclude the GPIb is tightly associated with ABP under conditions in which the endogenous CANP is inhibited, and that this apparent transmembrane complex of GPIb-ABP can be isolated in lysates of nonactivated human platelets.

Studies upon the Gram Reaction of the Basiphil Cells of the Anterior Pituitary

1952 ◽  
Vol s3-93 (22) ◽  
pp. 147-155
Author(s):  
C. L. FOSTER ◽  
R. R. WILSON
Keyword(s):  
Bile Salt ◽  
Positive Reaction ◽  
Anterior Pituitary ◽  
Periodic Acid ◽  
Point Of View ◽  
Methyl Green ◽  
Pituitary Cells ◽  
Periodic Acid Schiff ◽  
Human Anterior Pituitary ◽  
Micro Organisms

The basiphil cell granules of the human anterior pituitary react positively with Gram's stain, but no specific differentiation of these granules occurs when the iodine treatment is omitted. The granules of the acidophil cells are Gram-negative. Various workers have suggested a connexion between the Gram reaction of various micro-organisms and the presence of ribonucleoprotein, and the pituitary cells were investigated from this point of view. Pyronin methyl green staining demonstrates what is probably ribonucleoprotein material in the cytoplasm of both chromophobes and chromophils, but not in the granules of either kind of chromophil. As with micro-organisms, the Gram reaction is destroyed by treating the sections with hot oxygenated bile salt, which has a detergent action upon the ribonucleic acids, but, unlike micro-organisms, the reaction remains unaffected by digestion with a buffered solution of crystalline ribonuclease, although this treatment destroys the nucleic acid, as demonstrated by the loss of pyronin basiphilia in the cytoplasm. Bile salt also diminishes the strong positive reaction of the basiphil cell granules to the periodic acid Schiff (P.A.S.)test--a reaction which probably indicates the presence of intragranular mucoprotein. It seems unlikely that the Gramn-positive reaction of the basiphil cell granules of the human anterior pituitary is due simply to the presence of ribonucleoprotein. We have employed the term ribonucleic acid, bearing in mind that this is a generic term and not the name of a single molecular species (cf. Davidson, 1950).

scholarly journals Isolation and some structural analyses of a proteodermatan sulphate from calf skin

1983 ◽  
Vol 213 (2) ◽  
pp. 289-296 ◽  
Author(s):  
T Nakamura ◽  
E Matsunaga ◽  
H Shinkai
Keyword(s):  
Core Protein ◽  
Gradient Centrifugation ◽  
Cetylpyridinium Chloride ◽  
Periodic Acid ◽  
Molar Ratio ◽  
Gel Chromatography ◽  
Dermatan Sulphate ◽  
Periodic Acid Schiff ◽  
Cscl Density Gradient ◽  
Calf Skin

A proteodermatan sulphate was isolated from 0.15 M-NaCl and 0.45 M-NaCl extracts of newborn-calf skin. The proteoglycan was separated from collagen and hyaluronic acid by precipitation with cetylpyridinium chloride and CsCl-density-gradient centrifugation. Further purification was performed by ion-exchange, affinity and molecular-sieve chromatography. The proteoglycan bound to concanavalin A-Sepharose in 1 M-NaCl. It gave a positive reaction with periodic acid/Schiff reagent and contained 8.3% of uronic acid. The dermatan sulphate, the only glycosaminoglycan component, was composed of 74% iduronosylhexosamine units and 26% glucuronosylhexosamine units. The Mr was assessed to be 15000-20000 by gel chromatography. The core protein was found to be a sialoglycoprotein that had O-glycosidic oligosaccharides with N-acetylgalactosamine at the reducing termini. The molar ratio of oligosaccharide chains to dermatan sulphate was approx. 3:1. From these results the proposed structure of proteodermatan sulphate is: one dermatan sulphate chain (average Mr 17500), three O-glycosidic oligosaccharide chains and probably N-glycosidic oligosaccharide chain(s) bound to one core-protein molecule (Mr 55000).

scholarly journals Inhibition of RNA-binding protein HuR reduces glomerulosclerosis in experimental nephritis

2020 ◽  
Vol 134 (12) ◽  
pp. 1433-1448 ◽  
Author(s):  
Simeng Liu ◽  
Zhimin Huang ◽  
Anna Tang ◽  
Xiaoqing Wu ◽  
Jeffrey Aube ◽  
...  
Keyword(s):  
Rna Binding ◽  
Therapeutic Potential ◽  
Kidney Diseases ◽  
Binding Protein ◽  
Periodic Acid ◽  
Rna Binding Protein ◽  
Selective Inhibition ◽  
Control Group ◽  
Periodic Acid Schiff ◽  
Experimental Nephritis

Abstract Recent identification of an RNA-binding protein (HuR) that regulates mRNA turnover and translation of numerous transcripts via binding to an ARE in their 3′-UTR involved in inflammation and is abnormally elevated in varied kidney diseases offers a novel target for the treatment of renal inflammation and subsequent fibrosis. Thus, we hypothesized that treatment with a selective inhibition of HuR function with a small molecule, KH-3, would down-regulate HuR-targeted proinflammatory transcripts thereby improving glomerulosclerosis in experimental nephritis, where glomerular cellular HuR is elevated. Three experimental groups included normal and diseased rats treated with or without KH-3. Disease was induced by the monoclonal anti-Thy 1.1 antibody. KH-3 was given via daily intraperitoneal injection from day 1 after disease induction to day 5 at the dose of 50 mg/kg BW/day. At day 6, diseased animals treated with KH-3 showed significant reduction in glomerular HuR levels, proteinuria, podocyte injury determined by ameliorated podocyte loss and podocin expression, glomerular staining for periodic acid-Schiff positive extracellular matrix proteins, fibronectin and collagen IV and mRNA and protein levels of profibrotic markers, compared with untreated disease rats. KH-3 treatment also reduced disease-induced increases in renal TGFβ1 and PAI-1 transcripts. Additionally, a marked increase in renal NF-κB-p65, Nox4, and glomerular macrophage cell infiltration observed in disease control group was largely reversed by KH-3 treatment. These results strongly support our hypothesis that down-regulation of HuR function with KH-3 has therapeutic potential for reversing glomerulosclerosis by reducing abundance of pro-inflammatory transcripts and related inflammation.

scholarly journals Histogenesis of the Oesophagus of Guinea Fowl (Numida meleagris) at Prehatch and Posthatch

Scientifica ◽  
2016 ◽  
Vol 2016 ◽  
pp. 1-5
Author(s):  
Innocent Jonah Gosomji ◽  
Sulaiman Olawoye Salami ◽  
James Oliver Nzalak ◽  
Muhammed Umar Kawu ◽  
Emmanuel Vandi Tizhe ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Positive Reaction ◽  
Periodic Acid ◽  
Mesenchymal Cells ◽  
Guinea Fowl ◽  
Mucin Secretion ◽  
Periodic Acid Schiff ◽  
Numida Meleagris ◽  
Tunica Adventitia ◽  
Tunica Muscularis

The histogenesis of the primordial oesophagus was studied to determine the period in which the tunics of the oesophagus developed and became functional in the helmeted guinea fowl (Numida meleagris). Eighteen embryos and nine keets were studied at prehatch and posthatch, respectively. Simple columnar epithelium surrounded by mesenchymal cells was obvious at the 8th day of embryonic development. By the 19th day of embryonic development, the four tunics, tunica mucosa, submucosa, tunica muscularis, and tunica adventitia/serosa, were beginning to differentiate from the mesenchymal cells and also the primordial oesophageal glands appeared as clusters of cells that invaginate from the epithelium. By the 27th day, the tunics were clearly differentiated and the primordial glands were fully developed as evident with positive reaction to Periodic Acid Schiff (PAS). The tunics of the muscularis were not well developed till at posthatch. This study therefore concludes that the primordial oesophagus is active at the late incubation due to mucin secretion by mucous glands but fully functional at posthatch since the tunica muscularis is completely developed at posthatch.

The problem of Mehlis's gland in helminths with special reference to Penetrocephalus ganapatii (Cestoda: Pseudophyllidea)

Parasitology ◽  
1960 ◽  
Vol 50 (3-4) ◽  
pp. 349-350 ◽  
Author(s):  
K. Hanumantha-Rao
Keyword(s):  
Special Reference ◽  
Positive Reaction ◽  
Periodic Acid ◽  
Gland Secretion ◽  
Periodic Acid Schiff ◽  
Vitelline Cells ◽  
Egg Shell

1. Mehlis's gland secretion in trematodes and pseudophyllidean cestodes may be a lecithin-like phospholipid.2. After enzymic degradation of the lecithin into lysolecithin the release of egg-shell precursors from the vitelline cells is effected.3. Soon after the completion of the egg-shell the lysolecithin may be reconverted into lecithin in vitelline cells of Penetrocephalus ganapatii, thus accounting for the periodic acid-Schiff positive reaction.

scholarly journals Engineering and overexpression of periplasmic forms of the penicillin-binding protein 3 of Escherichia coli

1994 ◽  
Vol 298 (1) ◽  
pp. 189-195 ◽  
Author(s):  
C Fraipont ◽  
M Adam ◽  
M Nguyen-Distèche ◽  
W Keck ◽  
J Van Beeumen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Binding Protein ◽  
Molar Ratio ◽  
Amino Acid Residues ◽  
Penicillin Binding Protein ◽  
Operator Expression ◽  
Membrane Bound ◽  
Terminal Amino ◽  
Laci Repressor

Replacement of the 36 and 56 N-terminal amino acid residues of the 588-amino-acid-residue membrane-bound penicillin-binding protein 3 (PBP3) of Escherichia coli by the OmpA signal peptide allows export of F37-V577 PBP3 and G57-V577 PBP3 respectively into the periplasm. The modified ftsI genes were placed under the control of the fused lpp promoter and lac promoter/operator; expression of the truncated PBP3s was optimized by varying the copy number of the recombinant plasmids and the amount of LacI repressor, and export was facilitated by increasing the SecB content of the producing strain. The periplasmic PBP3s (yield 8 mg/l of culture) were purified to 70% protein homogeneity. They require the presence of 0.25 M NaCl to remain soluble. Like the membrane-bound PBP3, they undergo processing by elimination of the C-terminal decapeptide I578-S588, they bind penicillin in a 1:1 molar ratio and they catalyse hydrolysis and aminolysis of acyclic thioesters that are analogues of penicillin. The membrane-anchor-free PBP3s have ragged N-termini. The G57-V577 PBP3, however, is less prone to proteolytic degradation than the F37-V577 PBP3.

scholarly journals Purification and characterization of zinc-binding protein from the liver of the partially hepatectomized rat

1979 ◽  
Vol 183 (3) ◽  
pp. 683-690 ◽  
Author(s):  
H Ohtake ◽  
M Koga
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Binding Protein ◽  
Guanidine Hydrochloride ◽  
Deae Cellulose ◽  
Molar Ratio ◽  
Total Amino Acid ◽  
Amino Acid Residues

Zn-binding protein in liver of the partially hepatectomized rat was purified by column chromatography on Sephadex G-75 and DEAE-cellulose. Homogeneity was judged by polyacrylamide-disc-gel electrophoresis. The molecular weight determined by gel-permeation chromatography in 6 M-guanidine hydrochloride was 6700. This value is in good agreement with the molecular weight calculated from the amino acid composition, which was 6073. Zn-binding protein was composed of 61 amino acid residues, and the distinctive features include an extremely high content of cysteine, which accounted for one-third of the total amino acid residues, and an absolute absence of aromatic amino acids as well as of histidine, leucine and arginine. The amino acid composition was similar to that of the metallothioneins previously isolated from rat liver and mouse liver. These observations suggest that the Zn-binding protein can be classified as a type of metallothionein. Zn-binding protein contained 8.2g-atoms of zinc per mol and traces of copper, but no cadmium. The molar ratio of thiol groups to zinc was calculated to be 2.5:1. Possible roles of this Zn-binding protein in the transport and storage of zinc in the liver are discussed.

Ultrastructure of the Parotid Gland of the Nine-Banded Armadillo

1977 ◽  
Vol 35 ◽  
pp. 640-641
Author(s):  
J. R. Ruby
Keyword(s):  
Endoplasmic Reticulum ◽  
Parotid Gland ◽  
Periodic Acid ◽  
Acinar Cells ◽  
Duct System ◽  
Parotid Glands ◽  
Dasypus Novemcinctus ◽  
Anterior Portion ◽  
Periodic Acid Schiff ◽  
Thin Sections

Parotid glands were obtained from five adult (four male and one female) armadillos (Dasypus novemcinctus) which were perfusion-fixed. The glands were located in a position similar to that of most mammals. They extended interiorly to the anterior portion of the submandibular gland.In the light microscope, it was noted that the acini were relatively small and stained strongly positive with the periodic acid-Schiff (PAS) and alcian blue techniques, confirming the earlier results of Shackleford (1). Based on these qualities and other structural criteria, these cells have been classified as seromucous (2). The duct system was well developed. There were numerous intercalated ducts and intralobular striated ducts. The striated duct cells contained large amounts of PAS-positive substance.Thin sections revealed that the acinar cells were pyramidal in shape and contained a basally placed, slightly flattened nucleus (Fig. 1). The rough endoplasmic reticulum was also at the base of the cell.

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