Multiple molecular forms of avian aldolases. III. Tissue distribution and properties of the predominant isozymes from chicken (Gallus domesticus)

1969 ◽  
Vol 47 (12) ◽  
pp. 1187-1194 ◽  
Author(s):  
Ronald R. Marquardt
Keyword(s):  
Tissue Distribution ◽  
Heart Tissue ◽  
Isozyme Pattern ◽  
Gallus Domesticus ◽  
Molecular Forms ◽  
Molecular Weights ◽  
Aldolase C ◽  
Multiple Molecular Forms ◽  
Aldolase A

The distribution of chicken (Gallus domesticus) tissue (muscle, oviduct, heart, brain, kidney, and liver) aldolase isozymes was established electrophoretically. Each tissue, being composed of varying proportions of six isozymes, has a distinct isozyme pattern. Brain, oviduct, and heart tissue possess five isozymes of aldolase: 1, 2, 3, 4, and 5. Leg and breast muscle of adults possess aldolase 5. Kidney and liver predominantly possess aldolase 6 with traces of isozymes 1–5. Aldolase 1 corresponds to aldolase C of mammals, 5 to aldolase A, and 6 to aldolase B.Antibodies to 5 react with 2, 3, and 4, but not with 1 and 6. Aldolase 5 hybridizes in vitro with 1 to form 2, 3, and 4. Aldolase 6 also hybridizes in vitro with 1 to form three intermediate hybrids, but these hybrids have not been detected in tissues. These observations imply that aldolase 1 (C) and 5 (A) are frequently synthesized simultaneously in the cell with the formation of the 2, 3, and 4 hybrids. Aldolase 6 (B), in contrast, may not be accompanied by the synthesis of other aldolases in the same cell. Pure preparations of aldolases 1, 5, and 6 were found to have similar molecular weights but different enzymatic properties.

Multiple Molecular Forms of Avian Aldolases. V. Purification and Molecular Properties of Chicken (Gallus domesticus) Liver Aldolase

1971 ◽  
Vol 49 (6) ◽  
pp. 647-657 ◽  
Author(s):  
Ronald R. Marquardt
Keyword(s):  
Molecular Weight ◽  
Specific Activity ◽  
Diffusion Constant ◽  
Sedimentation Coefficient ◽  
Sedimentation Velocity ◽  
Gallus Domesticus ◽  
Molecular Forms ◽  
Velocity Analysis ◽  
Multiple Molecular Forms ◽  
Stokes Radius

Aldolase (fructose-1,6-diphosphate D-glyceraldehyde-3-phosphate-lyase, EC 4.1.2.13) was purified from chicken liver. The enzyme was shown to be homogeneous according to the following criteria: purification to a constant specific activity following sequential chromatography on carboxymethyl-Sephadex and Sephadex G-200, electrophoresis on cellulose acetate strips, sedimentation velocity analysis, absence of 10 other glycolytic enzymes, and immunodiffusion in agar.The sedimentation coefficient (s°20w 8.0), Stokes radius (47 Å), diffusion constant (D°20w 4.0 × 10−7 cm2/s), and molecular weight (160 000) are similar to those of rabbit liver aldolase and the muscle and brain enzymes from both chickens and rabbits.

Multiple molecular forms of avian aldolases. I. Crystallization and physical properties of chicken (Gallus domesticus) breast muscle aldolase

1969 ◽  
Vol 47 (5) ◽  
pp. 517-526 ◽  
Author(s):  
Ronald R. Marquardt
Keyword(s):  
Molecular Weight ◽  
Specific Activity ◽  
Diffusion Constant ◽  
Sedimentation Coefficient ◽  
Double Diffusion ◽  
Glycolytic Enzyme ◽  
Gallus Domesticus ◽  
Molecular Forms ◽  
Rabbit Muscle ◽  
Multiple Molecular Forms

Aldolase (fructose 1,6-diphosphate-D-glyceraldehyde 3-phosphate lyase, EC 4.1.2.13) was purified and crystallized from chicken (Gallus domesticus) breast muscle.The crystalline enzyme is homogeneous according to the following criteria: purification to a constant specific activity, electrophoresis on cellulose acetate strips, absence of five other glycolytic enzyme activities, and immunodiffusion in agar.The sedimentation coefficient, diffusion constant, and molecular weight of the chicken enzyme are the same as for rabbit muscle aldolase. The ultraviolet spectra of the two proteins are the same. Electrophoretic comparison between the rabbit and chicken enzymes revealed a slightly different rate of migration.Antibodies directed against the pure chicken enzyme were prepared, and the reaction with pure chicken and rabbit aldolase was followed using the precipitin and double diffusion tests. A very pronounced reaction was observed between anti-serum and the chicken enzyme; the rabbit enzyme, in contrast, did not cross-react with the anti-serum.

scholarly journals Multiple Molecular forms of Factor VIII Antigen in Normal Individuals and von Willebrand’s Disease Patients

1977 ◽  
Author(s):  
T. Zimmerman ◽  
J. Kimball ◽  
T. Edgington ◽  
C. Abildgaard
Keyword(s):  
Factor Viii ◽  
Molecular Forms ◽  
Type I ◽  
Type Ii ◽  
Von Willebrand's Disease ◽  
Von Willebrand’S Disease ◽  
Multiple Molecular Forms ◽  
Normal Individuals ◽  
Factor Viii Antigen

Factor VIII antigen is present in normal individuals in multiple molecular forms which can be separated according to size. The smaller forms have little or no ristocetin co-factor activity. In order to evaluate the possibility that Factor VIII antigen forms of large size may be an artifact of in vitro aggregation, we have ultracentrifuged plasma on a 20% sucrose cushion at 37 C for 10 min at 254,000 xg (peak). The rate of clearing of Factor VIII antigen was compared to that of other plasma proteins. The results indicate that Factor VIII-related antigen forms of high S exist even when plasma is maintained at physiological temperature and analyzed with minimal delay, suggesting that these larger molecular forms also exist as such in vivo.In von Willebrand’s disease (vWd) all forms of Factor VIII antigen may be present but decreased in quantity (Type I) or large forms may be missing with smaller forms present in normal or increased quantities (Type II). Factor VIII antigen was isolated from plasma of three patients with Type I and three patients with Type II vWd by counter immunoelectrophoresis. The Factor VIII antigen was then reduced and electrophoresed on SDS-containing Polyacrylamide gels. The presence of carbohydrate was evaluated by staining with perchloric acid-Schiff’s reagent (PAS). The 210,000 MW Factor VIII antigen subunit from each patient was PAS-positive. Though subtle changes in carbohydrate content or composition could not be evaluated by this technique, a total defect of glycosylation is unlikely in this sample of vWd patients.

Multiple molecular forms of prolactin during pregnancy in women

1985 ◽  
Vol 106 (1) ◽  
pp. 81-85 ◽  
Author(s):  
F. Pansini ◽  
C. M. Bergamini ◽  
M. Malfaccini ◽  
G. Cocilovo ◽  
M. Linciano ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Pregnant Women ◽  
Gradual Increase ◽  
Gel Filtration ◽  
Molecular Forms ◽  
Low Molecular Weight ◽  
Third Trimester ◽  
Molecular Weights ◽  
The Third ◽  
Multiple Molecular Forms

ABSTRACT The distribution of isomorphic forms of prolactin in the serum of pregnant women was studied by gel filtration chromatography. Using this technique we were able to resolve three peaks, detected by radioimmunoassay: they were termed 'big-big', 'big' and 'little' prolactin in order of decreasing size, with approximate molecular weights > 100 000, 50 000 and 21 000 respectively. They displayed a comparable immunoreactivity to the antiserum employed in the radioimmunoassay, as determined in competition experiments. The relative amount of each hormone form in serum changed during the third trimester of pregnancy. At week 33 of pregnancy, 'little' prolactin accounted for 63·2 ± 7·7% of the total circulating hormone present in the serum of five normal pregnant women. During the progression of pregnancy, there was a gradual increase in the low molecular weight prolactin, so that, at the time of delivery, the larger forms of the hormone were present only in small amounts. J. Endocr. (1985) 106, 81–85

Soluble Thrombomodulin Purified from Human Urine Exhibits a Potent Anticoagulant Effect In Vitro and In Vivo

1995 ◽  
Vol 73 (05) ◽  
pp. 805-811 ◽  
Author(s):  
Yasuo Takahashi ◽  
Yoshitaka Hosaka ◽  
Hiromi Niina ◽  
Katsuaki Nagasawa ◽  
Masaaki Naotsuka ◽  
...  
Keyword(s):  
Human Urine ◽  
Specific Activity ◽  
Anticoagulant Activity ◽  
Rabbit Lung ◽  
Soluble Thrombomodulin ◽  
Molecular Weights ◽  
Dose Dependent Fashion ◽  
Dose Dependent

SummaryWe examined the anticoagulant activity of two major molecules of soluble thrombomodulin purified from human urine. The apparent molecular weights of these urinary thrombomodulins (UTMs) were 72,000 and 79,000, respectively. Both UTMs showed more potent cofactor activity for protein C activation [specific activity >5,000 thrombomodulin units (TMU)/mg] than human placental thrombomodulin (2,180 TMU/mg) and rabbit lung thrombomodulin (1,980 TMU/mg). The UTMs prolonged thrombin-induced fibrinogen clotting time (>1 TMU/ml), APTT (>5 TMU/ml), TT (>5 TMU/ml) and PT (>40 TMU/ml) in a dose-dependent fashion. These effects appeared in the concentration range of soluble thrombomodulins present in human plasma and urine. In the rat DIC model induced by thromboplastin, administration of UTMs by infusion (300-3,000 TMU/kg) restored the hematological abnormalities derived from DIC in a dose-dependent fashion. These results demonstrate that UTMs exhibit potent anticoagulant and antithrombotic activities, and could play a physiologically important role in microcirculation.

Toxicity of Heparinoids, with Special Reference to the Precipitation of Fibrinogen

1964 ◽  
Vol 12 (01) ◽  
pp. 232-261 ◽  
Author(s):  
S Sasaki ◽  
T Takemoto ◽  
S Oka
Keyword(s):  
Molecular Weight ◽  
Dextran Sulfate ◽  
Anticoagulant Activity ◽  
Molecular Structures ◽  
Severe Reaction ◽  
Clinical Use ◽  
Molecular Weights ◽  
Close Relationship

SummaryTo demonstrate whether the intravascular precipitation of fibrinogen is responsible for the toxicity of heparinoid, the relation between the toxicity of heparinoid in vivo and the precipitation of fibrinogen in vitro was investigated, using dextran sulfate of various molecular weights and various heparinoids.1. There are close relationships between the molecular weight of dextran sulfate, its toxicity, and the quantity of fibrinogen precipitated.2. The close relationship between the toxicity and the precipitation of fibrinogen found for dextran sulfate holds good for other heparinoids regardless of their molecular structures.3. Histological findings suggest strongly that the pathological changes produced with dextran sulfate are caused primarily by the intravascular precipitates with occlusion of the capillaries.From these facts, it is concluded that the precipitates of fibrinogen with heparinoid may be the cause or at least the major cause of the toxicity of heparinoid.4. The most suitable molecular weight of dextran sulfate for clinical use was found to be 5,300 ~ 6,700, from the maximum value of the product (LD50 · Anticoagulant activity). This product (LD50 · Anticoagulant activity) can be employed generally to assess the comparative merits of various heparinoids.5. Clinical use of the dextran sulfate prepared on this basis gave satisfactory results. No severe reaction was observed. However, two delayed reactions, alopecia and thrombocytopenia, were observed. These two reactions seem to come from the cause other than intravascular precipitation.

Platelet Microtubule Subunit Proteins

1979 ◽  
Vol 42 (05) ◽  
pp. 1630-1633 ◽  
Author(s):  
A G Castle ◽  
N Crawford
Keyword(s):  
Epithelial Cells ◽  
Gel Electrophoresis ◽  
Blood Platelets ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Lens Epithelial Cells ◽  
Molecular Weights ◽  
Kinetics Of ◽  
Microtubular Structures

SummaryBlood platelets contain microtubule proteins (tubulin and HMWs) which can be polymerised “in vitro” to form structures which resemble the microtubules seen in the intact platelet. Platelet tubulin is composed of two non-identical subunits a and p tubulin which have molecular weights around 55,000 but can be resolved in alkaline SDS-polyacrylamide gel electrophoresis. These subunits associate as dimers with sedimentation coefficients of about 5.7 S although it is not known whether the dimer protein is a homo- or hetero-dimer. The dimer tubulin binds the anti-mitotic drug colchicine and the kinetics of this binding are similar to those reported for neurotubulins. Platelet microtubules also contain two HMW proteins which appear to be essential and integral components of the fully assembled microtubule. These proteins have molecular weights greater than 200,000 daltons. Fluorescent labelled antibodies to platelet and brain tubulins stain long filamentous microtubular structures in bovine lens epithelial cells and this pattern of staining is prevented by exposing the cells to conditions known to cause depolymerisation of cell microtubules.

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