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.

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
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Sea Urchin ◽  
Cellular Localization ◽  
Binding Protein ◽  
Gel Permeation Chromatography ◽  
Total Amino Acid ◽  
Amino Acid Residues ◽  
Gel Permeation ◽  
Anthocidaris Crassispina

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.

Preparation and fractionation of goat κ-casein: analysis of the glycan and peptide components

1978 ◽  
Vol 45 (2) ◽  
pp. 191-196 ◽  
Author(s):  
Francesco Addeo ◽  
Solange Soulier ◽  
Jean-Pierre Pelissier ◽  
Jean-Marc Chobert ◽  
Jean-Claude Mercier ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Deae Cellulose ◽  
Phosphate Content ◽  
Carbohydrate Composition ◽  
Neuraminic Acids ◽  
Main Fraction ◽  
Carbohydrate Contents

SummaryWhole goat κ-casein was prepared by chromatography of whole casein on hydroxyapatite. Chromatography of whole κ-casein on DEAE-cellulose separated 5 fractions. All of them were sensitive to chymosin. Their amino acid and carbohydrate composition, phosphate content and molecular weight were determined. Galactose, N-acetylgalactosamine, N-acetyl and N-glycolyl neuraminic acids were identified in whole κ-casein. It appears that goat κ-casein, like cow, buffalo and ewe κ-caseins, is composed of several fractions having identical peptide chains and differing in their carbohydrate contents. The main fraction, devoid of carbohydrate, was treated with chymosin. The para-κ-casein and caseinomacropeptide were isolated. Their amino acid composition and phosphate content were determined.

ORNITHO-KININOGEN: ITS ISOLATION AND CHARACTERIZATION

1987 ◽  
Author(s):  
M Kimura ◽  
T Sueyoshi ◽  
T Morita ◽  
K Tanaka ◽  
S Iwanaga
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Reversed Phase ◽  
Amino Acid Residues ◽  
Isolation And Characterization ◽  
Bovine Plasma ◽  
Sds Page ◽  
Plasma Kinins

In mammals, three kinds of kininogens, high-Mr kininogen, low-Mr kininogen and T-kininogen, are known and their structures and functions have been well studied. However, little is known about non-mammalian kininogens and their existence has been known only by the release of plasma kinins. In 1966, Werle et al. reported that an ornitho-kinin liberated by tissue kallikrein from chicken, plasma differs;from mammalian kinins in its amino acid composition. Since that, there is no report about the structure of ornitho-kinin.In this study, ornitho-kininogen was isolated from chicken plasma by two steps with S-alkylated papain affinity and DEAE-5PW columns. The yield was 1.7 mg from 44 ml of plasma. The purified material gave a single band on SDS-PAGE with or without 2-mercaptoethanoL and on disc-PAGE, and the molecular weight was estimated to be 74,000 on SDS-PAGE by Ferguson plot. Ornitho-kininogen seemed to belong to mammalian high-Mr kininogen, based on the amino acid composition, and the molecular weight, and no kininogen corresponding to low-Mr kininogen and T kininogen was found in chicken plasma. In fact, ornitho-kininogen was degraded rapidly by bovine plasma kallikrein, liberating an ornitho-kinin, and the resulting kinin was separated by reversed phase HPLC on a column of Cosmosil 5C18-P. The amino acid sequence of ornitho-kinin was established to be Arg-Pro-Pro-Gly-Phe-Thr-Pro-Leu-Arg, in which the composition was different from that reported earlier by Werle et al. This sequence was similar to that of bradykinin but the two substitutions of Thr-6 and Leu-8 for Ser and Phe were found. Ornitho-kinin induced a hypotension on chicken and contracted the isolated smooth muscle. However, it did not any effect on the isolated rat uterus, suggesting that the specificity may be due to replacement of Phe-8 by Leu. The NH2-terminal 30 amino acid residues of ornitho-kininogen exhibited 43 % identity with that of bovine kininogen.

Hog pancreatic α-amylase. Preparation and characterization

1979 ◽  
Vol 44 (1) ◽  
pp. 288-293 ◽  
Author(s):  
Ivan Kluh
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Disulfide Bonds ◽  
Polypeptide Chain ◽  
Deae Cellulose ◽  
Amino Acid Residues ◽  
Single Polypeptide Chain ◽  
Single Polypeptide ◽  
End Group

Crystalline α-amylase (EC 3.2.1.1) was prepared from hog pancreas. The preparation obtained was resolved into two isozymes by chromatography on DEAE-cellulose. The molecular weight (51 500), amino acid composition, and terminal groups of both isozymes were determined. Both isozymes have a single polypeptide chain containing 460-465 amino acid residues. The amino acid composition of both isozymes is similar. None of them has a free N-terminal end group. Both isozymes are C-terminated with leucine. The molecule of each isozyme is cross-linked by 5 disulfide bonds and contains two sulfhydryl groups.

scholarly journals The primary structure of aspartate aminotransferase from pig heart muscle. Partial sequences determined by digestion with thermolysin and elastase

1973 ◽  
Vol 133 (4) ◽  
pp. 805-819 ◽  
Author(s):  
Francesco Bossa ◽  
Donatella Barra ◽  
Massimo Carloni ◽  
Paolo Fasella ◽  
Francesca Riva ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Primary Structure ◽  
Aspartate Aminotransferase ◽  
Heart Muscle ◽  
Science And Technology ◽  
Amino Acid Sequences ◽  
Amino Acid Residues ◽  
Lending Library

Peptides produced by thermolytic digestion of aminoethylated aspartate aminotransferase and of the oxidized enzyme were isolated and their amino acid sequences determined. Digestion by elastase of the carboxymethylated enzyme gave peptides representing approximately 40% of the primary structure. Fragments from these digests overlapped with previously reported sequences of peptides obtained by peptic and tryptic digestion (Doonan et al., 1972), giving ten composite peptides containing 395 amino acid residues. The amino acid composition of these composite peptides agrees well with that of the intact enzyme. Confirmatory results for some of the present data have been deposited as Supplementary Publication 50018 at the National Lending Library for Science and Technology, Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1973) 131, 5.

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