scholarly journals Sea urchin (Anthocidaris crassispina) egg zinc-binding protein. Cellular localization, purification and characterization

1983 ◽  
Vol 211 (1) ◽  
pp. 109-118 ◽  
Author(s):  
H Ohtake ◽  
T Suyemitsu ◽  
M Koga

Gel-filtration analysis of cytosol fraction obtained from unfertilized sea-urchin (Anthocidaris crassispina) eggs on Sephadex G-75 revealed the presence of two Zn-binding-protein fractions. The major Zn-binding protein fraction had a low molecular weight and a low absorbance at 280 nm, properties similar to those of the metallothionein found in the regenerating rat liver. These fractions were further purified by DEAE-cellulose and Sephadex G-50 chromatography. Homogeneity of the Zn-binding protein was judged by polyacrylamide-disc-gel electrophoresis and gel-permeation chromatography in the presence of 6 M-guanidinium chloride. The molecular weight determined by gel-permeation chromatography was 3900. This value is in good agreement with the minimum molecular weight calculated from the amino acid composition, which was 3655. Zn-binding protein is composed of 36 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 a complete absence of aromatic amino acids, as well as of methionine, histidine and arginine. Zn-binding protein contained 4.1 g-atoms of zinc per mol and a trace of cadmium, but no copper, iron or calcium. The molar ratio of reactive thiol groups to metal ion was calculated to be 2.73:1. Possible roles of this Zn-binding protein in the homoeostasis of zinc in unfertilized sea-urchin eggs are discussed.

1979 ◽  
Vol 183 (3) ◽  
pp. 683-690 ◽  
Author(s):  
H Ohtake ◽  
M Koga

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.


1980 ◽  
Vol 192 (2) ◽  
pp. 569-577 ◽  
Author(s):  
J Barr ◽  
P Nordin

Membrane preparations from Acer pseudoplatanus suspension cultures were demonstrated to incorporate radioactivity from GDP-[U-14C]mannose and UDP-N-acetyl-[6-(3)H]glucosamine into high-molecular-weight polymers characterized as glycoprotein. From 20 to 25% of the 14C was incorporated as fucose with the remainder as mannose, whereas 90% of the 3H was incorporated as N-acetylglucosamine with the remainder as N-acetylgalactosamine. Pronase digestion yielded radioactive glycopeptides that were separated into four fractions by gel-permeation chromatography and paper electrophoresis. The isolated glycopeptides differed in molecular weight and isotopes incorporated, as well as in amino-acid and monosaccharide composition. The membrane preparation also incorporated radioactivity from the added nucleotides into chloroform/methanol (2:1, v/v)- and chloroform/methanol/water (10:10:3, by vol.)-soluble lipids, and into an insoluble pellet.


2012 ◽  
Vol 487 ◽  
pp. 48-52
Author(s):  
Sheng Hua Lv ◽  
Gong Rui ◽  
Di Li

The radical copolymerization of resorcinol (RSC) and 3,5-dihydroxyl benzoic (DHBA) was carried out in water by the initiator of horseradish (HRP)/H2O2. It was discussed that the effects of monomer composition on the properties of the copolymer. The best monomer mass ratio of RSC:DHBA was 60:40 and the shrink temperature can reached to 88.5oC. The tanning result indicated that the copolymer of RSC and DHPA has particularly excellent tanning properties and can be served as leather tannage substitute for chrome tanning materials. And also the mechanism of the HRP initiated copolymerization was proposed. The structure and molecular weight of the copolymer was characterized by Fourier Transform Infrared spectroscopy (FTIR), Nuclear Magnetic Resonance (NMR) and Gel Permeation Chromatography (GPC). The copolymer can be as tannage and retannage in making leather process. The results showed that it has excellent tanning properties and retanning effects.


1995 ◽  
Vol 311 (2) ◽  
pp. 407-415 ◽  
Author(s):  
W C Buhi ◽  
I M Alvarez ◽  
V M Shille ◽  
M J Thatcher ◽  
J P Harney ◽  
...  

A major canine endometrial secreted protein (cP6, 23,000-M(r)) was purified by ion-exchange and gel-filtration chromatography and characterized by two-dimensional gel electrophoresis. Anti-[human retinol-binding protein (hRBP)] serum identified cP6 on immunoblot analysis and immunoprecipitated cP6 from culture medium. This major protein was also shown to bind [3H]retinol. N-terminal and internal amino acid sequences were determined and compared with previously identified protein, RNA, or DNA sequences. N-terminal analysis revealed that cP6 had high identity and similarity to serum retinol-binding proteins (RBPs), while internal sequence analysis showed a strong similarity to rat androgen-dependent epididymal protein and beta-lactoglobulins. Amino acid analysis, however, showed significant differences between these proteins and cP6 in both total amino acid content and certain selected amino acids. Immunohistochemical analysis showed staining for RBP only in the uterine luminal epithelium. These studies suggest that bitch endometrium secretes a family of proteins (cP6), some of which bind [3H]retinol, are immunologically related to the RBP family, and have N-terminal and internal sequences with a high similarity to RBP, beta-lactoglobulins and other members of the lipocalin family. This family of proteins may be important in early development for supplying retinol or derivatives to the developing embryo.


1975 ◽  
Vol 28 (1) ◽  
pp. 189 ◽  
Author(s):  
RA Shanks

Gel permeation columns of Bio Beads S-X8 have been used to provide separation of oligomers and other small organic molecules. Results show successful separations up to molecular weight c. 600. The retention times of compounds have been correlated with the largest molecular dimension of the molecules and also with molar volumes.


Author(s):  
Cigdem Kilicarislan Ozkan ◽  
Hasan Ozgunay

Dialdehyde starches with different aldehyde content from native corn starch were prepared by sodium periodate oxidation to be used as a tanning agent in leather making. For this purpose, native corn starch was oxidized with sodium metaperiodate in different molar ratios. After oxidation processes, the yields, solubility in water and aldehyde contents of the obtained dialdehyde starches were determined as well as structure characterizations by Proton Nuclear Magnetic Resonance Spectroscopy, Fourier Transform Infrared Spectroscopy and Gel Permeation Chromatography. Evaluating the gel permeation chromatography data, the dialdehyde starch samples which were thought to be in appropriate molecular weight/size to penetrate into skin fibers were selected to be used in the tanning process. Their tanning abilities were evaluated by investigating hydrothermal stabilities, filling and fiber isolation characteristics and physical properties determined by mechanical tests and organoleptically. From the evaluation of the results, it was revealed that sodium metaperiodate oxidized starches which have appropriate molecular weight and adequate aldehyde content has a remarkable tanning effect and can be utilized as a tanning agent with the advantages of not necessitating pickling process which means saving time and simplifying the production but more importantly offering an important advantage from an environmental point of view.


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