THE HETEROGENEITY OF GAMMA-GLOBULIN IN POST-EXERCISE URINE

1964 ◽  
Vol 42 (7) ◽  
pp. 1065-1097 ◽  
Author(s):  
M. H. Freedman ◽  
G. E. Connell
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Polypeptide Chain ◽  
Gel Filtration ◽  
Low Molecular Weight ◽  
Starch Gel Electrophoresis ◽  
Single Individual ◽  
Post Exercise ◽  
Γ Globulins ◽  
Normal Urine

Post-exercise urine was collected and the protein was precipitated with ammonium sulphate. The γ-globulin was separated from other urinary proteins by preparative starch block electrophoresis. The γ-globulin was then separated by gel filtration into two fractions, the faster one consisting mainly of 7 S γ-globulin while the slower one contained "low molecular weight" γ-globulin sedimenting at 3.5 S. The low molecular weight γ-globulins were further fractionated on CM-Sephadex to yield four fractions. Each of the chromatographic fractions was shown to be heterogeneous by starch gel electrophoresis at pH 4. The same degree of heterogeneity was observed in preparations of pooled urinary γ-globulin and γ-globulin from a single individual. The two major electrophoretic components of each chromatographic fraction formed precipitates in agar diffusion tests with antiserum to 7 S γ-globulin and to the L-polypeptide chain of 7 S γ-globulin. Two chromatographic fractions reacted with antiserum to the H-polypeptide chain of 7 S γ-globulin. All of the fractions failed to react with antiserum to β2A-globulin and β2M-globulin. Amino acid analysis showed distinct differences among the chromatographic fractions. One of the fractions closely resembled the L-chain of 7 S plasma γ-globulin with respect to amino acid composition. After reduction of disulphide bonds and alkylation most of the urinary γ-globulin resembles L-chain in electrophoretic behavior. The γ-globulins of post-exercise urine were found to be qualitatively similar to the γ-globulins of normal urine but post-exercise γ-globulins were present quantitatively in much larger amounts.

scholarly journals The isolation of three neurophysins from porcine posterior pituitary lobes

1970 ◽  
Vol 116 (5) ◽  
pp. 899-909 ◽  
Author(s):  
L. O. Uttenthal ◽  
D. B. Hope
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Analysis ◽  
Gel Filtration ◽  
Biological Significance ◽  
Starch Gel Electrophoresis ◽  
Posterior Pituitary ◽  
Fresh Tissue ◽  
Molecular Weights ◽  
Molecular Dimensions

1. Three neurophysins, proteins that bind the polypeptide hormones oxytocin and vasopressin, have been isolated from acetone-dried porcine posterior pituitary lobes. The proteins have been named porcine neurophysins-I, -II and -III in order of their electrophoretic mobilities at pH8.1. 2. Electrophoretic comparison of the purified proteins, which are homogeneous on starch-gel electrophoresis, with the soluble proteins of fresh porcine posterior pituitary lobes extracted in 0.1m-HCl and in buffer pH8.1 suggests that the isolated proteins are native to the fresh tissue. 3. Neurophysins-I and -II are present in similar amounts in the tissue, whereas neurophysin-III is present only in small quantities. Acetone-dried tissue also contains traces of other hormone-binding neurophysin components. 4. All the neurophysins can bind both oxytocin and [8-lysine]-vasopressin. 5. The apparent molecular weights of the neurophysins increase with increasing protein concentration as measured by equilibrium sedimentation in the ultracentrifuge. 6. Neurophysins-I and -III are of similar molecular dimensions, contain one residue of methionine per molecule and lack histidine. The minimum molecular weight of neurophysin-I obtained by amino acid analysis is 9360. Neurophysin-II is of larger molecular dimensions than neurophysins-I and -III and can be separated from these by gel filtration on Sephadex G-75. It contains no histidine or methionine, and its minimum molecular weight has been estimated as 14020 by amino acid analysis. 7. Each of the three neurophysins possesses N-terminal alanine. 8. The possible biological significance of the existence of several neurophysins within one species is discussed.

Limited Proteolysis of the Purified Aα Chain of Human Fibrinosen by Plasmin

1975 ◽  
Author(s):  
L. Williams ◽  
G. Murano
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Peptide Mapping ◽  
Proteolytic Enzymes ◽  
Degradation Products ◽  
Limited Proteolysis ◽  
Gel Filtration ◽  
Terminal Region ◽  
Low Molecular Weight ◽  
Human Fibrinogen

Based on evidence that a portion of circulating fibrinogen consists of a family of catabolic intermediates formed by proteolytic degradation of the COOH terminal region of Aα chains, we attempted to obtain early degradation products using the purified alkylated Aα chain derivative of human fibrinogen as the substrate and plasmin as the enzyme. Having established optimal conditions, a preparative quantity of material was digested in 0.1 M tris buffer pH = 9.5; time = 4 min; E/S ratio = 1/75 (mole/mole); temp = 37° C. Low molecular weight fragments were separated from the larger species, and further purified by gel filtration on Sephadex G-100. Selected early fragments were analyzed by polycrylamide gel electrophoresis, amino acid composition, peptide mapping and partial N-terminal amino acid sequence. Two of the earliest low molecular weight fragments released by plasmin were derived from the N-terminal region of the Aα chain. Their molecular size was estimated at about 10,000 daltons. One fragment contains fibrinopeptide A; both fragments extend beyond Met-51. Our data indicate that: a) the specificity of plasmin on the purified Aα chain differs from that on intact fibrinogen; or b) proteolytic enzymes other than or in addition to plasmin are responsible for the formation of early catabolic fibrinogen intermediates having a degraded Aα chain.(Supported by USPHS N. I. H. Grant HL 14142.)

The Nature of the Chelate Structures Formed by Copper(II) and Ovalbumin in the Biuret Reaction

1963 ◽  
Vol 16 (6) ◽  
pp. 989 ◽  
Author(s):  
AC Jennings
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Alkaline Solution ◽  
Polypeptide Chain ◽  
Chemical Reactivity ◽  
Spectrophotometric Study ◽  
Low Molecular Weight ◽  
Amino Acid Residues ◽  
Heterogeneous Mixture

A spectrophotometric study was made of the interaction, in alkaline solution, of copper(II) with ovalbumin and several ligands of low molecular weight at several different atomic ratios of nitrogen/copper. The results indicate that the absorbance and chemical reactivity of the ovalbumin-copper(II) complex cannot be attributed to the presence of one particular chelate structure. It is suggested that the ovalbumin-copper(II) complex is a heterogeneous mixture of chelate structures in which the copper(II) is bonded to oxygen (in the enolate form) and nitrogen atoms in the several different combinations possible within the same polypeptide chain. It is also suggested that the actual arrangement of bonds in any one chelate structure is probably determined by both the nature of the amino acid residues and the configuration of the ovalbumin molecule in the immediate vicinity.

MONOMER AND DIMER FORMS OF GAMMA-GLOBULIN POLYPEPTIDES IN NORMAL URINE

1964 ◽  
Vol 42 (12) ◽  
pp. 1815-1823 ◽  
Author(s):  
M. H. Freedman ◽  
G. E. Connell
Keyword(s):  
Molecular Weight ◽  
Disulfide Bonds ◽  
Gamma Globulin ◽  
The Other ◽  
Light Chains ◽  
Low Molecular Weight ◽  
Γ Globulins ◽  
Intermolecular Disulfide Bonds ◽  
Normal Urine ◽  
Polypeptide Chains

The γ-globulins of low molecular weight in human urine have been partially fractionated and characterized. The two principal components are made up of "light" polypeptide chains of 7 S γ-globulin. One of these components, the "monomer", has a molecular weight in dissociating solvents of approximately 20,000 and probably consists of single light chains. The other component, the "dimer", has a molecular weight of approximately 40,000 and probably consists of pairs of light chains joined by intermolecular disulfide bonds.

In Vivo Studies on the Inhibition of Coagulation by Fractionated Heparin and by a Heparin Analogue I. Effects of Heparin Fractions

1981 ◽  
Vol 46 (03) ◽  
pp. 612-616 ◽  
Author(s):  
U Schmitz-Huebner ◽  
L Balleisen ◽  
F Asbeck ◽  
J van de Loo
Keyword(s):  
Molecular Weight ◽  
Day Treatment ◽  
Gel Filtration ◽  
Weight Fraction ◽  
In Vivo Studies ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
Platelet Factor ◽  
Heparin Fractions

SummaryHigh and low molecular weight heparin fractions obtained by gel filtration chromatography of sodium mucosal heparin were injected subcutaneously into six healthy volunteers and compared with the unfractionated substance in a cross-over trial. Equal doses of 5,000 U were administered twice daily over a period of three days and heparin activity was repeatedly controlled before and 2, 4, 8 hrs after injection by means of the APTT, the anti-Xa clotting test and a chromogenic substrate assay. In addition, the in vivo effect of subcutaneously administered fractionated heparin on platelet function was examined on three of the volunteers. The results show that s.c. injections of the low molecular weight fraction induced markedly higher anti-Xa activity than injections of the other preparations. At the same time, APTT results did not significantly differ. Unfractionated heparin and the high molecular weight fraction enhanced ADP-induced platelet aggregation and collagen-mediated MDA production, while the low molecular weight fraction hardly affected these assays, but potently inhibited thrombin-induced MDA production. All heparin preparations stimulated the release of platelet Factor 4 in plasma. During the three-day treatment periods, no side-effects and no significant changes in the response to heparin injections were detected.

scholarly journals The complete amino acid sequence of the low molecular weight cytosolic acid phosphatase

1989 ◽  
Vol 264 (5) ◽  
pp. 2560-2567
Author(s):  
G Camici ◽  
G Manao ◽  
G Cappugi ◽  
A Modesti ◽  
M Stefani ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Acid Phosphatase ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Low Molecular Weight ◽  
Complete Amino Acid Sequence

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 Characterization of Trypsin Inhibitor in Florunner Peanut Seeds (Arachis hypogaea L.)1

1988 ◽  
Vol 15 (2) ◽  
pp. 81-84 ◽  
Author(s):  
E. M. Ahmed ◽  
J. A. Applewhite
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Aspartic Acid ◽  
Arachis Hypogaea ◽  
Trypsin Inhibitor ◽  
Low Molecular Weight ◽  
Arachis Hypogaea L ◽  
Amino Acid Profiles ◽  
Low Levels

Abstract Florunner peanut seeds contained five trypsin isoinhibitors. Amino acid profiles of the trypsin inhibitors fraction showed high levels of aspartic acid, half-cystine and serine and low levels of histidine and tyrosine. The molecular weight of the inhibitor was 8.3 KDa. The presence of multiforms of this inhibitor, its low molecular weight and the high amount of half-cystine indicate that peanut trypsin inhibitor is of the Bowman-Birk type.

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.

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