Antifreeze glycoproteins in the plasma of Newfoundland Atlantic cod (Gadus morhua)

1981 ◽  
Vol 59 (11) ◽  
pp. 2186-2192 ◽  
Author(s):  
Choy L. Hew ◽  
Don Slaughter ◽  
Garth L. Fletcher ◽  
Shashikant B. Joshi
Keyword(s):  
Amino Acid ◽  
High Performance ◽  
Gadus Morhua ◽  
Atlantic Cod ◽  
Antifreeze Proteins ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Polar Cod ◽  
Antifreeze Glycoproteins ◽  
Molecular Weights

The plasma of the Atlantic cod, Gadus morhua, contained antifreeze glycoproteins which were present only during the winter months. The antifreeze proteins were isolated, using gel filtration and ion exchange chromatography, and characterized by high performance liquid chromatography. The antifreeze proteins appeared to consist of at least seven components with molecular weights ranging from 2 500 to 33 000. Chemical analysis of the larger components showed a predominance of alanine, threonine, and galactosamine in a ratio of 2:1:1. The smaller peptides contained proline, in addition to alanine and threonine. The amino acid sequence of the smallest glycopeptide (molecular weight 2500) was found to be Ala Ala Thr Pro Ala Thr Ala Ala Thr Pro Ala Thr Ala Ala.These glycoproteins are very similar, if not identical, in amino acid and carbohydrate composition to those isolated from Antaractic nototheniids and several northern gadoids. The sequence of the smallest glycopeptide from the Atlantic cod is identical to that reported for the polar cod, Boreogadus saida.

Isolation and characterization of antifreeze glycoproteins from the frostfish, Microgadus tomcod

1982 ◽  
Vol 60 (3) ◽  
pp. 348-355 ◽  
Author(s):  
Garth L. Fletcher ◽  
Choy L. Hew ◽  
Shashikant B. Joshi
Keyword(s):  
Amino Acid ◽  
High Performance ◽  
Antifreeze Proteins ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Antifreeze Glycoproteins ◽  
Molecular Weights ◽  
Isolation And Characterization ◽  
Saffron Cod ◽  
Microgadus Tomcod

The antifreeze proteins were isolated from frostfish (Microgadus tomcod) using gel filtration and ion exchange chromatography and characterized by high-performance liquid chromatography. The antifreeze proteins were glycoproteins which appeared to consist of at least six components with molecular weights ranging from 2 550 to 32 200. Chemical analysis of the larger components showed a predominance of alanine, threonine, and galactosamine. The smaller peptides contained proline and arginine in addition to alanine and threonine. The amino acid sequence of the smallest glycopeptides (molecular weight 2 550) was found to be Ala-Ala-Thr-Ala-Ala-Thr-[Formula: see text]-Ala-Thr-Ala-Ala-Thr-Pro-Ala-[Formula: see text]-Ala-Ala.These glycopeptides are very similar in amino acid and carbohydrate composition to those isolated from the Antarctic nototheniids and several northern gadoids. The sequence of the first 14 amino acids of smaller glycopeptides from the frostfish is identical to comparable peptides found in the nototheniids and the saffron cod. To date arginine has only been observed in the glycopeptide antifreezes of the frostfish and the saffron cod.

Production of antifreeze glycoproteins in cultured and wild juvenile Atlantic cod (Gadus morhua L.) in a common laboratory environment

2001 ◽  
Vol 79 (4) ◽  
pp. 610-615 ◽  
Author(s):  
C F Purchase ◽  
S V Goddard ◽  
J A Brown
Keyword(s):  
High Temperatures ◽  
Gadus Morhua ◽  
Gulf Of Maine ◽  
Atlantic Cod ◽  
Antifreeze Proteins ◽  
Antifreeze Glycoproteins ◽  
Laboratory Environment ◽  
Grand Banks ◽  
Common Laboratory ◽  
Northeast Coast

Many fishes accumulate antifreeze proteins or antifreeze glycoproteins (AFGPs) in the blood to increase their chances of survival in cold seawater. Cod (Gadus morhua L.) from colder environments have been found to produce more AFGPs than those from warmer areas, but the genetic and environmental contributions to this variation have not been determined. Populations of cultured (from the Grand Banks; Gulf of Maine) and wild (from Fortune Bay; Bonavista Bay) juvenile cod were kept in a common laboratory environment to investigate differences in AFGP production. All the populations were capable of producing AFGPs, and the AFGP levels were similar in cultured and wild cod. The results indicate that high temperatures associated with the production of cultured cod do not negatively affect the ability to produce AFGPs. In addition, young cod from as far south as the Gulf of Maine are capable of producing AFGPs at levels similar to those from the northeast coast of Newfoundland.

Isolation and partial characterization of boar proacrosin

1990 ◽  
Vol 55 (3) ◽  
pp. 846-853 ◽  
Author(s):  
Věra Jonáková ◽  
Brigita Vidimská ◽  
Jana Urbanová ◽  
Manfred Pavlík
Keyword(s):  
Amino Acid ◽  
High Performance ◽  
Gel Filtration ◽  
Reversed Phase ◽  
Exchange Chromatography ◽  
Relative Molecular Mass ◽  
Reducing Conditions ◽  
Apparent Homogeneity ◽  
Step Procedure

Boar proacrosin was purified to apparent homogeneity by a three-step procedure: gel filtration on Sephadex G-50 medium, ion-exchange chromatography on CM-Cellulose 32, and reversed-phase high-performance liquid chromatography on a C4 column. The relative molecular mass (Mr) of the proacrosin estimated by gel filtration was about 70 000, whereas the results of an electrophoretic experiment on SDS-polyacrylamide gel with copolymerized casein under non-reducing conditions indicated an Mr of 55 000-60 000. The proacrosin reproducibly migrated on the gel as a double band. When purified, it remained stable at pH 8.0 for 30 min. The amino-acid composition of the homogeneous proacrosin was determined, the N-terminal amino-acid sequence being Arg-Asp-X-Ala-Thr-X-X-Gly-Pro-X-Gly-.

Preparation and Characterization of NH2-Terminal Fibrinogen Bβ Fragments from N-DSK of Human Fibrinogen

1984 ◽  
Vol 51 (01) ◽  
pp. 016-021 ◽  
Author(s):  
S Birken ◽  
G Agosto ◽  
B Lahiri ◽  
R Canfield
Keyword(s):  
High Performance ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Reverse Phase ◽  
Human Fibrinogen ◽  
Human Volunteers ◽  
Cnbr Cleavage ◽  
Two Peaks

SummaryIn order to investigate the early release of NH2-terminal plasmic fragments from the Bβ chain of fibrinogen, substantial quantities of Bβ 1-42 and Bβ 1-21 are required as immunogens, as radioimmunoassay standards and for infusion into human volunteers to determine the half-lives of these peptides. Towards this end methods that employ selective proteolytic cleavage of these fragments from fibrinogen have been developed. Both the N-DSK fragment, produced by CNBr cleavage of fibrinogen, and Bβ 1-118 were employed as substrates for plasmin with the finding of higher yields from N-DSK. Bβ 1-42 and Bβ 1-21 were purified by gel filtration and ion-exchange chromatography on SP-Sephadex using volatile buffers. When the purified preparation of Bβ 1-42 was chromatographed on reverse-phase high performance liquid chromatography, two peaks of identical amino acid composition were separated, presumably due either to pyroglutamate or to amide differences.

Bovine Factors X1And X2: Activation Peptide Based Chromatographic Differences

1979 ◽  
Author(s):  
Takashi Morita ◽  
Craig Jackson
Keyword(s):  
Amino Acid ◽  
Anion Exchange ◽  
Gel Filtration ◽  
Heart Association ◽  
Personal Communication ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Factor X ◽  
Activation Peptide ◽  
Activation Peptides

Bovine Factor X is eluted in two forms (X1and X2) from anion exchange chromatographic columns. These two forms have indistinguishable amino acid compositions, molecular weights and specific activities. The amino acid sequences containing the γ-carboxyglutamic acid residues have been shown to be identical in X1 and X2(H. Morris, personal communication). An activation peptide is released from the N-terminal region of the heavy chain of Factor X by an activator from Russell’s viper venom. This peptide can be isolated after activation by gel filtration on Sephadex G-100 under nondenaturing conditions. The activation peptides from a mixture of Factors X1 and X2 were separated into two forms by anion-exchange chromatography. The activation peptide (AP1) which eluted first was shown to be derived from Factor X1. while the activation peptiae (AP2) which eluted second was shown to be derived from X2 on the basis of chromatographic separations carried out on Factors X1 and X2 separately. Factor Xa was eluted as a symmetrical single peak. On the basis of these and other data characterizing these products, we conclude that the difference between X1 and X2 are properties of the structures of the activation peptides. (Supported by a grant HL 12820 from the National Heart, Lung and Blood Institute. C.M.J. is an Established Investigator of the American Heart Association).

Co-purification of bone resorbing activity and adenylate cyclase stimulating activity from human tumours associated with the humoral hypercalcaemia of malignancy

1987 ◽  
Vol 114 (1) ◽  
pp. 18-26 ◽  
Author(s):  
Chohei Shigeno ◽  
Itsuo Yamamoto ◽  
Shegiharu Dokoh ◽  
Megumu Hino ◽  
Jun Aoki ◽  
...  
Keyword(s):  
Bone Resorption ◽  
High Performance ◽  
Basic Protein ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Protein Factor ◽  
Human Tumours ◽  
Acid Stable ◽  
Bone Resorbing Activity

Abstract. We have partially purified a tumour factor capable of stimulating both bone resorption in vitro and cAMP accumulation in osteoblastic ROS 17/2 cells from three human tumours associated with humoral hypercalcaemia of malignancy. Purification of tumour factor by sequential acid urea extraction, gel filtration and cation-exchange chromatography, reverse-phase high performance liquid chromatography followed by analytical isoelectric focussing provided a basic protein (pI > 9.3) with a molecular weight of approximately 13 000 as a major component of the final preparation which retained both the two bioactivities. Bone resorbing activity and cAMP-increasing activity in purified factor correlated with each other. cAMP-increasing activity of the factor was heat- and acid-stable, but sensitive to alkaline ambient pH. Treatment with trypsin destroyed cAMP-increasing activity of the factor. Synthetic parathyroid hormone (PTH) antagonist, human PTH-(3– 34) completely inhibited the cAMP-increasing activity of the factor. The results suggest that this protein factor, having its effects on both osteoclastic and osteoblastic functions, may be involved in development of enhanced bone resorption in some patients with humoral hypercalcaemia of malignancy.

scholarly journals Chemical identity of tryptensin with angiotensin

1980 ◽  
Vol 187 (3) ◽  
pp. 647-653 ◽  
Author(s):  
K Arakawa ◽  
M Yuki ◽  
M Ikeda
Keyword(s):  
Amino Acid ◽  
Thin Layer ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Human Plasma Protein ◽  
Plasma Protein Fraction ◽  
Layer Chromatography

Tryptensin, a vasopressor substance generated from human plasma protein fraction IV-4 by trypsin, has been isolated and the amino acid composition analysed. The procedures used for the isolation were: (a) adsorption of the formed tryptensin on Dowex 50W (X2; NH4+ form); (b) gel filtration through Sephadex G-25; (c) cation-exchange chromatography on CM-cellulose; (d) anion-exchange chromatography on DEAE-cellulose; (e) re-chromatography on CM-cellulose; (f) gel filtration on Bio-Gel P-2; (g) partition chromatography on high-pressure liquid chromatography. The homogeneity of the isolated tryptensin was confirmed by thin-layer chromatography and thin-layer electrophoresis. The amino acid analysis of the hydrolysate suggested the following proportional composition: Asp, 1; Val, 1; Ile, 1; Tyr, 1; Phe, 1; His, 1; Arg, 1; Pro, 1. This composition is identical with that of human angiotensin.

Effect of extractant and molecular size on the optical and chemical properties of soil humic acids

Soil Research ◽  
1969 ◽  
Vol 7 (3) ◽  
pp. 229 ◽  
Author(s):  
JHA Butler ◽  
JN Ladd
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Humic Acids ◽  
Gel Filtration ◽  
Chemical Properties ◽  
Molecular Size ◽  
Carboxyl Content ◽  
Peptide Bonds ◽  
Molecular Weights ◽  
Amino Acid Contents

Humic acids extracted from soil with sodium pyrophosphate have greater proportions of lower molecular weight material, less acid-hydrolysable amino acid nitrogen contents, but greater carboxyl contents and extinction values (260 and 450 nm) than humic acids extracted subsequently from the same sample with alkali. Humic acids extracted with alkali from fresh soil samples have intermediate values. Extinction values at 260 nm are directly correlated with carboxyl contents for a given soil. Different crop histories have no significant effect on the measured properties of the extracted humic acids. An alkali-extracted humic acid has been fractionated by gel filtration into seven fractions of different nominal molecular weight ranges. As the molecular weights of the fractions increase, both aliphatic C-H (based on infrared absorption at 2900 cm-1) and acid-hydrolysable amino acid contents increase, whereas extinction values at 260 nm and carboxyl contents decrease. The infrared spectra of the high molecular weight fractions have peaks at 1650 and 1510 cm-1 which correlate with acid-hydrolysable amino acid contents and which correspond to amide I and II bands of peptide bonds. Alkaline hydrolysis to split peptide bonds eliminates both these peaks. The spectra also have peaks at 1720 and 1210 cm-1 which correlate with the carboxyl content.

scholarly journals α-Crystallin. The isolation and characterization of distinct macromolecular fractions

1971 ◽  
Vol 124 (2) ◽  
pp. 337-343 ◽  
Author(s):  
Abraham Spector ◽  
Lu-Ku Li ◽  
Robert C. Augusteyn ◽  
Arthur Schneider ◽  
Thomas Freund
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Chemical Difference ◽  
Molecular Weights ◽  
Isolation And Characterization ◽  
Different Populations ◽  
Molecular Weight Fractions

α-Crystallin was isolated from calf lens periphery by chromatography on DEAE-cellulose and gel filtration. Three distinct populations of macromolecules have been isolated with molecular weights in the ranges approx. 6×105−9×105, 0.9×106−4×106and greater than 10×106. The concentration of macromolecules at the molecular-weight limits of a population are very low. The members of the different populations do not appear to be in equilibrium with each other. Further, in those molecular-weight fractions investigated, no equilibrium between members of the same population was observed. The population of lowest molecular weight comprises 65–75% of the total material. The amino acid and subunit composition of the different-sized fractions appear very similar, if not identical. The only chemical difference observed between the fractions is the presence of significant amounts of sugar in the higher-molecular-weight fractions. Subunit molecular weights of approx. 19.5×103and 22.5×103were observed for all α-crystallin fractions.

scholarly journals The Fine Structure of the Sensory Epithelium of the Vomeronasal Organ in Suckling Rats

1971 ◽  
Vol 24 (3) ◽  
pp. 765 ◽  
Author(s):  
Jean E Kratzing
Keyword(s):  
Amino Acid ◽  
Gel Filtration ◽  
Vomeronasal Organ ◽  
Sensory Epithelium ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Tryptic Digestion ◽  
Tryptic Peptides ◽  
Special Procedure ◽  
A Chain

The amino acid sequence of the a-chain of haemoglobin from M. giganteus has been determined. The soluble peptides formed by tryptic digestion were isolated by gel filtration, ion-exchange chromatography, paper ionophoresis, and chromatography. The amino acid sequences were determined by the "dansyl"Edman procedure. Incomplete hydrolysis of one bond resulted in a large insolublecore peptide containing 40 amino acid residues. The sequence of this peptide was deduced from the sequences of smaller peptides resulting from further digestion with thermolysin and papain. Maleylation of the a-globin before tryptic digestion gave three large fragments which assisted in assigning tryptic peptides to specific areas of the molecule. A special procedure involving maleylation of a chymotryptic digest of globin was used to isolate peptides containing arginine which provided overlap sequences of tryptic peptides

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