scholarly journals Chemical and physical characterization of a proline-rich polypeptide from sheep colostrum

1981 ◽  
Vol 199 (1) ◽  
pp. 9-15 ◽  
Author(s):  
M Janusz ◽  
K Starościk ◽  
M Zimecki ◽  
Z Wieczorek ◽  
J Lisowski
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Room Temperature ◽  
Proteolytic Enzymes ◽  
Gel Filtration ◽  
Guanidinium Chloride ◽  
Terminal Amino Acid ◽  
Polyproline Ii ◽  
Terminal Amino

A proline-rich polypeptide isolated from sheep colostrum is described. The molecular weight of the polypeptide determined by gel filtration is 17 200. However, in the presence of guanidinium chloride the molecular weight found is about 6000. The polypeptide contains about 22% of proline, a high proportion of non-polar amino acids, a low percentage of glycine, and no alanine, arginine and cysteine residues. The only N-terminal amino acid found is leucine. C.d. spectra in water and in 50% (v/v) trifluoroethanol suggest the presence of block sequences of proline residues forming helices of polyproline II type. The proline-rich polypeptide is soluble at 4 degrees C but is reversibly precipitated on warming to room temperature. Maximal precipitation is observed at pH 4.6 and at ionic strength above 0.6. The precipitation depends on the concentration of the polypeptide. No effect of other proteins, Ca2+ and Zn2+ ions on the precipitation of the polypeptide was found. The proline-rich polypeptide is not an amphipathic protein. The lack of effect of the polypeptide on proteolytic enzymes ruled out the possibility that it is an inhibitor of proteinases.

scholarly journals Characterization of gastric mucoproteins isolated by equilibrium density-gradient centrifugation in caesium chloride

1974 ◽  
Vol 141 (3) ◽  
pp. 633-639 ◽  
Author(s):  
Bryan J. Starkey ◽  
David Snary ◽  
Adrian Allen
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Gradient Centrifugation ◽  
Gel Filtration ◽  
Equilibrium Density ◽  
Amino Acid Residues ◽  
Gastric Mucus ◽  
Terminal Amino Acid ◽  
Terminal Amino

1. The mucoprotein from pig gastric mucus has been purified by equilibrium centrifugation in a CsCl gradient. 2. This procedure removes the non-covalently bound protein, which is closely associated with the mucoprotein and not easily removed from it by gel filtration. 3. The purified mucoprotein is separable by gel filtration into a high-molecular-weight mucoprotein A (mol.wt. 2.3×106) and a low-molecular-weight mucoprotein B/C (mol.wt. 1.15×106). 4. These two mucoproteins have the same chemical analysis namely fucose 11.3%, galactose 26%, glucosamine 19.5%, galactosamine 8.3% and protein 13.6%. 5. Mucoprotein A contains 3.1% ester sulphate. 6. These mucoproteins are isolated without enzymic digestion and have a higher protein content than the blood-group-substance mucoproteins from proteolytic digestion of gastric mucus. Detailed amino acid analysis shows that the extra protein in the non-enzymically digested material is composed of amino acids other than serine and threonine. 7. Mucoproteins A and B/C contain respectively 130 and 9 half-cystine residues per molecule of which about 78 and 6 residues are involved in disulphide linkages. 8. Cleavage of these disulphide linkages by mercaptoethanol splits both mucoproteins into four equally sized subunits of mol.wt. 5.2×105for mucoprotein A and 2.8×104for mucoprotein B/C. 9. The sole N-terminal amino acid of mucoprotein A is aspartic acid, whereas mucoprotein B/C has several different N-terminal amino acid residues.

CHARACTERIZATION OF THE PIGMENT OF MICROCOCCUS VIOLAGABRIELLAE

1964 ◽  
Vol 10 (5) ◽  
pp. 659-675 ◽  
Author(s):  
J. N. Campbell ◽  
J. L. Nichols ◽  
Sheila A. Berry
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Cross Reactivity ◽  
Low Molecular Weight ◽  
Terminal Amino Acid ◽  
Iron Chelate ◽  
Peptide Moiety ◽  
Terminal Amino

Production of the red insoluble pigment by Micrococcus violagabriellae was studied. Pigmentation was found to require oxygen and high levels of iron and to be stimulated by tryptophan alone among the amino acids. The pigment was isolated, purified, and analyzed chemically and spectrophotometrically. It was found to be similar to pulcherrimin from Candida pulcherrima. Immunological cross reactivity and analysis of derivatives confirmed the similarity between the bacterial and yeast pigments. From these data it is postulated that the pigment is an iron chelate of pulcherriminic acid with an associated low molecular weight peptide moiety with glycine as the sole N-terminal amino acid. The pigment appears to differ from that of C. pulcherrima solely with respect to this peptide and in the mode of aggregation of the molecule.

Characterization of an isozyme of S-adenosylmethionine synthetase from rat liver

1984 ◽  
Vol 62 (5) ◽  
pp. 276-279 ◽  
Author(s):  
C. H. Lin ◽  
W. Chung ◽  
K. P. Strickland ◽  
A. J. Hudson
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Enzymatic Synthesis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Aromatic Amino Acids ◽  
Adenosylmethionine Synthetase ◽  
Cellulose Column

An isozyme of S-adenosylmethionine synthetase has been purified to homogeneity by ammonium sulfate fractionation, DEAE-cellulose column chromatography, and gel filtration on a Sephadex G-200 column. The purified enzyme is very unstable and has a molecular weight of 120 000 consisting of two identical subunits. Amino acid analysis on the purified enzyme showed glycine, glutamate, and aspartate to be the most abundant and the aromatic amino acids to be the least abundant. It possesses tripolyphosphatase activity which can be stimulated five to six times by S-adenosylmethionine (20–40 μM). The findings support the conclusion that an enzyme-bound tripolyphosphate is an obligatory intermediate in the enzymatic synthesis of S-adenosylmethionine from ATP and methionine.

scholarly journals Rabbit Tamm–Horsfall urinary glycoprotein. Chemical composition and subunit structure

1971 ◽  
Vol 122 (5) ◽  
pp. 623-631 ◽  
Author(s):  
Anne M. S. Marr ◽  
A. Neuberger ◽  
Wendy A. Ratcliffe
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Chemical Composition ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Cross Reactivity ◽  
Subunit Structure ◽  
Terminal Amino

1. Tamm–Horsfall glycoprotein from rabbit urine has been isolated and characterized. The homogeneity of the preparation has been established by a variety of procedures including disc gel electrophoresis and ultracentrifugation in aqueous solution, sodium dodecyl sulphate and formic acid. 2. The chemical composition has been determined and a carbohydrate content of approx. 31% was obtained. The relative contents of the amino acids were shown to be very similar to those in human Tamm–Horsfall glycoprotein. A trace of lipid was also detected. 3. Leucine was identified as the only N-terminal amino acid. 4. The subunit structure was investigated in the presence of sodium dodecyl sulphate by gel filtration and disc gel electrophoresis. These studies indicated that the subunit possessed a molecular weight of approx. 84000±6000. A similar value was obtained after reduction and S-alkylation of the glycoprotein indicating that the disulphide bonds were all intrachain. 5. A minimum value for the chemical molecular weight of 85000±6000 was obtained from the number of N-terminal amino acids released by cyanogen bromide cleavage of the glycoprotein. 6. The immunological properties of the glycoprotein were studied. Cross reactivity was demonstrated between human Tamm–Horsfall glycoprotein and a guinea-pig anti-rabbit Tamm–Horsfall antiserum.

Isolation and characterization of β-lipolytic hormone from porcine pituitary gland

1970 ◽  
Vol 48 (9) ◽  
pp. 1017-1021 ◽  
Author(s):  
C. Gilardeau ◽  
M. Chrétien
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Pituitary Gland ◽  
Amino Acid Composition ◽  
Biological Activities ◽  
Terminal Amino Acid ◽  
Isolation And Characterization ◽  
Pituitary Glands ◽  
Terminal Amino

A lipolytic substance was isolated from porcine pituitary glands. It's amino acid composition, molecular weight, N-terminal amino acid, isoelectric point, and biological activities are reported. These results are compared to the corresponding values of sheep β-lipolytic hormone.

Isolation and characterization of a zinc-containing metalloprotease expressed by Vibrio tubiashii

2003 ◽  
Vol 49 (8) ◽  
pp. 525-529 ◽  
Author(s):  
R B Delston ◽  
M H Kothary ◽  
K A Shangraw ◽  
B D Tall
Keyword(s):  
Gel Filtration ◽  
Hydrophobic Interaction Chromatography ◽  
Ammonium Sulfate Precipitation ◽  
Terminal Amino Acid ◽  
Isolation And Characterization ◽  
Vibrio Tubiashii ◽  
Strong Homology ◽  
Isoelectric Points ◽  
Terminal Amino

A Vibrio tubiashii hemagglutinin, a protease, was purified by ammonium sulfate precipitation, gel filtration, and hydrophobic interaction chromatography. It agglutinates sheep, chicken, bovine, rabbit, guinea pig, and human erythrocytes. It has a molecular mass of 35 kDa, isoelectric points of 3.5 and 3.7, and is inhibited by ortho-phenanthro line, phosphoramidon, and Zincov. The N-terminal amino acid sequence (Ala-Gln-Ala-Thr-Gly-Thr-Gly- Pro-Gly-Gly-Asn-Gln-Lys-Thr-Gly-Gln- Tyr-Asn-Phe-Gly) has strong homology to other Vibrio proteases.Key words: Vibrio tubiashii, metalloprotease, hemagglutinin.

scholarly journals Isolation and characterization of a 30 kDa protein with antifungal activity from leaves of Engelmannia pinnatifida

1996 ◽  
Vol 316 (3) ◽  
pp. 723-727 ◽  
Author(s):  
Q. Khai HUYNH ◽  
Jeffry R. BORGMEYER ◽  
Christine E. SMITH ◽  
Leslie D. BELL ◽  
Dilip M. SHAH
Keyword(s):  
Antifungal Activity ◽  
Plant Pathogens ◽  
Gel Filtration ◽  
Self Incompatibility ◽  
Terminal Amino Acid ◽  
Reverse Phase ◽  
Ammonium Sulphate Precipitation ◽  
Isolation And Characterization ◽  
Terminal Amino

During the course of screening plants for novel antifungal activity, we found that a high-molecular-mass fraction of an extract from leaves of Engelmannia pinnatifida exhibited potent and broad-spectrum antifungal activity. In this study a 30 kDa protein from E. pinnatifida leaves was purified to homogeneity by ammonium sulphate precipitation, gel filtration, Mono-Q and C18 reverse-phase column chromatographies. The purified protein showed potent antifungal activity against various plant pathogens with as little as 50 ng. The N-terminal amino acid sequence of the purified protein was determined as XXTKFDFFTLALQXPAXF, where X indicates an unidentified residue. This sequence showed 35–50% sequence identity with purified style glycoproteins associated with self-incompatibility from wild tomato, tobacco and petunia, a phosphate-starvation-induced ribonuclease from cultured tomato cells and the SIR 63.4 kDa protein from yeast.

scholarly journals Preparation and characterization of fragments from the N-terminal end of bovine serum albumin under native conditions

1993 ◽  
Vol 296 (2) ◽  
pp. 347-350 ◽  
Author(s):  
T Saeed ◽  
A Salahuddin
Keyword(s):  
Amino Acid ◽  
Gel Filtration ◽  
Proteolytic Digestion ◽  
Thiol Groups ◽  
Optical Studies ◽  
Terminal Amino Acid ◽  
Hydrodynamic Parameters ◽  
Terminal Amino ◽  
Domain I

The domain I of BSA, containing residues 1-183 of the protein sequence, was isolated by CNBr treatment. It was further reductively cleaved into two subfragments, N1 and N2, in 8 M urea; the subfragments were regenerated in GSH and GSSG. The fragment N and subfragments N1 and N2 were found to be homogeneous with respect to size and charge. Results for amino acid composition, N-terminal amino acid sequence, thiol groups and M(r) suggested that the fragments N1 and N2 contain residues 88-183 and 1-87 of the intact BSA respectively. Optical studies, intrinsic-viscosity measurements, gel-filtration data and derived hydrodynamic parameters, taken together with the results on proteolytic digestion, showed that fragment N, as well as its subfragments N1 and N2, exist in compact and globular conformation and that the conformation of N2 fragment is more compact than that of the N1 fragment.

CHARACTERIZATION OF AMINO ACID AND CARBOHYDRATE COMPONENTS IN FULVIC ACID

1976 ◽  
Vol 56 (3) ◽  
pp. 159-166 ◽  
Author(s):  
G. GUIDI ◽  
G. PETRUZZELLI ◽  
P. SEQUI
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Fulvic Acid ◽  
Weight Distribution ◽  
Gel Filtration ◽  
Percentage Content ◽  
Acidic Amino Acids ◽  
Carbohydrate Components

The distribution of individual amino acids and monosaccharides in fulvic acid and its fractions separated by polyamide chromatography was investigated in five different Italian soils. Although little differences were generally found in the two polyamide fractions (FI and FII), the highest percentage content of acidic amino acids and the lowest percentage content of neutral amino acids have been found in the second one (FII); monosaccharides composition was more irregular, but generally FII contained more pentoses. Both chromatographic fractions (FI and FII) have been chromatographed on Sephadex G-25. The composition in carbohydrate and amino acid components of the further different fractions resolved by gel filtration showed great differences depending on the molecular weight distribution.

IMPROVED PROCEDURES FOR PREPARATION AND CHARACTERIZATION OF MYROTHECIUM CELLULASE: PART 3. MOLECULAR WEIGHT, AMINO ACID COMPOSITION, TERMINAL RESIDUES, AND OTHER PROPERTIES

1963 ◽  
Vol 41 (3) ◽  
pp. 697-705 ◽  
Author(s):  
P. K. Datta ◽  
K. R. Hanson ◽  
D. R. Whitaker
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Edman Degradation ◽  
Terminal Amino Acid ◽  
Fingerprint Pattern ◽  
Sulphydryl Groups ◽  
Terminal Amino

The molecular weight of Myrothecium cellulase was estimated by the Archibald method to be approximately 49,000. No N-terminal amino acid could be detected by the Edman degradation or with fluorodinitrobenzene. Hydrazinolysis gave glycine as the C-terminal amino acid. No free sulphydryl groups could be detected in the enzyme. The amino acid composition and the fingerprint pattern after tryptic digestion were determined.

Sign in / Sign up

Export Citation Format

Share Document