scholarly journals The organ-specificity of ferritin in human and horse liver and spleen

1973 ◽  
Vol 131 (1) ◽  
pp. 51-59 ◽  
Author(s):  
R. R. Crichton ◽  
J. A. Millar ◽  
R. L. C. Cumming ◽  
C. F. A. Bryce
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Organ Specificity ◽  
Subunit Molecular Weight ◽  
Electrophoretic Mobilities ◽  
A Value ◽  
Horse Liver ◽  
Normal Human Liver ◽  
Quaternary Structures ◽  
Species Specific

1. Ferritin was isolated from human and horse spleen and liver, and apoferritin prepared therefrom. 2. The electrophoretic mobilities of the four apoferritins were determined on polyacrylamide gels and on cellulose acetate strips, and all found to be equal. 3. Homologous ferritins share reactions of identity in immunodiffusion experiments, whereas heterologous ferritins show only partial identity. 4. The subunit molecular weight of each of the apoferritins was determined by polyacrylamide-gel electrophoresis in sodium dodecyl sulphate and by chromatography on agarose columns in 6m-guanidine–HCl. A value of approx. 18500 was found in all cases. The proteins all had sedimentation coefficients of 17–18S. It thus seems that they have identical quaternary structures. 5. The amino acid compositions of the proteins revealed distinct differences both between organs and between species. This was confirmed by analysis of the tryptic peptide patterns, where it was found that about one-third of the peptides were common to the four proteins and the other two-thirds varied from protein to protein. 6. It is concluded that the apoferritins present in the liver and spleen of human and horse are both organ- and species-specific. 7. The apoferritin isolated from the liver of a patient with idiopathic haemochromatosis was identical with normal human liver apoferritin by the criteria described above.

Novel Catalatic Proteins of Bakers' Yeast. I. An Atypical Catalase

1973 ◽  
Vol 51 (11) ◽  
pp. 1551-1555 ◽  
Author(s):  
Tony C. M. Seah ◽  
A. R. Bhatti ◽  
J. G. Kaplan
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Native Protein ◽  
Wild Type ◽  
Major Band ◽  
Subunit Molecular Weight ◽  
Purification And Properties

At any stage of growth of a wild-type bakers' yeast, some 20% of the catalatic activity of crude extracts is not precipitable by means of antibody prepared against the typical catalase (catalase T), whose purification and properties have been previously described. Some of this catalatic activity is due to the presence of an atypical catalase (catalase A), a heme protein, with a molecular weight estimated as 170 000 – 190 000, considerably lower than that of the usual catalases (225 000 – 250 000). Preparations of catalase A were found to be homogeneous in the analytical ultracentrifuge and in polyacrylamide gel electrophoresis. Its subunit molecular weight, determined from its iron content, was 46 500, virtually the same as that of the major band obtained in gel electrophoresis in the presence of sodium dodecyl sulfate, suggesting that the native protein is tetrameric. Its specific activity is in the range of those reported for other typical catalases.

scholarly journals Increased membrane binding of erythrocyte catalase in hereditary spherocytosis and in metabolically stressed normal cells

Blood ◽  
1977 ◽  
Vol 49 (1) ◽  
pp. 113-123 ◽  
Author(s):  
DW Allen ◽  
S Cadman ◽  
SR McCann ◽  
B Finkel
Keyword(s):  
Molecular Weight ◽  
Hereditary Spherocytosis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Membrane Binding ◽  
Sodium Dodecyl ◽  
Protein Band ◽  
Normal Cells ◽  
Calcium Dependent ◽  
Subunit Molecular Weight ◽  
Erythrocyte Catalase

Abstract Normal red blood cell (RBC) membranes were compared with (1) RBC membranes from six patients with hereditary spherocytosis (HS), (2) normal membranes after hemolysis of the RBC in the presence of calcium, or (3) normal membranes after incubation of RBC for 24 hr in phosphate- buffered saline containing calcium without added glucose. When compared with normal controls, polyacrylamide gel electrophoresis with sodium dodecyl sulfate (PAGE SDS) of all three preparations showed an increase in membrane binding of globin and protein band 4.5 (60,000 molecular weight). In an attempt to identify band 4.5, 14 enzymes were assayed in the RBC membranes. Of these, catalase and lactate dehydrogenase were increased in membranes from HS RBC and from normal cells exposed to calcium. Only catalase, however, was present in sufficient quantity and had the correct subunit molecular weight on PAGE SDS and calcium- dependent membrane binding to account for an appreciable portion of 4.5. Caralase was further identified with a component of band 4.5 by double immunodiffusion using a specific anti-catalase antibody.

scholarly journals Selective depletion of small basic non-glycosylated proteins in diabetes

1981 ◽  
Vol 198 (1) ◽  
pp. 149-157 ◽  
Author(s):  
F C Samaniego ◽  
F Berry ◽  
J F Dice
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Liver Cytosol ◽  
Physiological Importance ◽  
Subunit Molecular Weight ◽  
Uncontrolled Diabetes ◽  
Streptozotocin Induced Diabetes ◽  
Protein Classes ◽  
Rat Liver Cytosol ◽  
Glycosylated Proteins

Degradative rates of small basic non-glycosylated proteins are preferentially enhanced in rat liver cytosol during severe streptozotocin-induced diabetes. Synthetic rates of these classes of proteins are not selectively enhanced in diabetes, so small basic non-glycosylated proteins should be depleted from liver cytosol as a consequence of this disease. To test this hypothesis, proteins were analysed from normal animals, from diabetic animals receiving insulin and from diabetic animals after insulin withdrawal for 3 days. The proteins were separated according to subunit molecular weight by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, according to isoelectric point by isoelectric focusing and according to carbohydrate content by affinity chromatography with concanavalin A linked to agarose. Severe uncontrolled diabetes is associated with the predicted depletion of small basic non-glycosylated proteins from liver cytosol. The preferential degradation and loss of these protein classes may be of considerable physiological importance to the animal.

Studies on the Dissociation of Porcine Haptoglobin

1972 ◽  
Vol 50 (7) ◽  
pp. 775-781 ◽  
Author(s):  
W. L. Lockhart ◽  
W. P. Chung ◽  
David B. Smith
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Light Chains ◽  
Dilute Solution ◽  
Disulfide Bridges ◽  
Elution Volume ◽  
Reversible Dissociation ◽  
Electrophoretic Mobilities ◽  
Chain Composition

Porcine haptoglobin was tested for reversible dissociation in dilute solution by gel filtration. Elution volume in Bio-Gel P-150 was independent of concentration and shapes of elution profiles failed to show dissociation. Molecular weight in dilute solution was estimated as 96 500. Interchain disulfide bridges were assayed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Partially reduced samples showed a series of intermediates but fully reduced samples showed only heavy and light chains. Intermediates were tentatively identified as to chain composition from their electrophoretic mobilities.

scholarly journals Cloning and Expression of a DNA Sequence Encoding a 41-Kilodalton Cryptosporidium parvum Oocyst Wall Protein

1999 ◽  
Vol 6 (6) ◽  
pp. 912-920 ◽  
Author(s):  
Mark C. Jenkins ◽  
Jim Trout ◽  
Charles Murphy ◽  
James A. Harp ◽  
Jim Higgins ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Recombinant Dna ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cloning And Expression ◽  
Wall Material ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Oocyst Wall ◽  
Species Specific

ABSTRACT This study was conducted to produce a recombinant species-specific oocyst wall protein of Cryptosporidium parvum. Antigens unique to C. parvum were identified by gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of oocyst proteins from several different Cryptosporidiumspecies. Antiserum was then prepared against a 41-kDa antigen unique toC. parvum and used to identify a recombinant DNA clone, designated rCP41. Expression of CP41 mRNA in C. parvumoocysts was confirmed by reverse transcriptase PCR (RT-PCR). Although the CP41 sequence was shown by PCR to be present in the genome ofC. baileyi, CP41 mRNA was not detected in this species by RT-PCR. Immunofluorescence staining with antiserum against recombinant CP41 detected native CP41 antigen on the surface of C. parvum oocysts but failed to detect CP41 on C. baileyi oocysts. Immunoelectron microscopy demonstrated that native CP41 was distributed unevenly on the C. parvumoocyst surface and was associated with amorphous oocyst wall material. In an enzyme-linked immunosorbent assay, purified rCP41 performed as well as native C. parvum oocyst protein in measuring the serological responses of young calves and adult cows to experimental and natural C. parvum infections. These results indicate that recombinant CP41 antigen may have potential in the immunodiagnosis of cryptosporidiosis.

scholarly journals Increased membrane binding of erythrocyte catalase in hereditary spherocytosis and in metabolically stressed normal cells

Blood ◽  
1977 ◽  
Vol 49 (1) ◽  
pp. 113-123
Author(s):  
DW Allen ◽  
S Cadman ◽  
SR McCann ◽  
B Finkel
Keyword(s):  
Molecular Weight ◽  
Hereditary Spherocytosis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Membrane Binding ◽  
Sodium Dodecyl ◽  
Protein Band ◽  
Normal Cells ◽  
Calcium Dependent ◽  
Subunit Molecular Weight ◽  
Erythrocyte Catalase

Normal red blood cell (RBC) membranes were compared with (1) RBC membranes from six patients with hereditary spherocytosis (HS), (2) normal membranes after hemolysis of the RBC in the presence of calcium, or (3) normal membranes after incubation of RBC for 24 hr in phosphate- buffered saline containing calcium without added glucose. When compared with normal controls, polyacrylamide gel electrophoresis with sodium dodecyl sulfate (PAGE SDS) of all three preparations showed an increase in membrane binding of globin and protein band 4.5 (60,000 molecular weight). In an attempt to identify band 4.5, 14 enzymes were assayed in the RBC membranes. Of these, catalase and lactate dehydrogenase were increased in membranes from HS RBC and from normal cells exposed to calcium. Only catalase, however, was present in sufficient quantity and had the correct subunit molecular weight on PAGE SDS and calcium- dependent membrane binding to account for an appreciable portion of 4.5. Caralase was further identified with a component of band 4.5 by double immunodiffusion using a specific anti-catalase antibody.

Plasma and Platelet Fibrinogen Differ in γ Chain Content

1984 ◽  
Vol 51 (01) ◽  
pp. 084-088 ◽  
Author(s):  
Charles W Francis ◽  
Ralph L Nachman ◽  
Victor J Marder
Keyword(s):  
Polypeptide Chain ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Human Platelets ◽  
Immunoaffinity Chromatography ◽  
Plasma Fibrinogen ◽  
Western Blots ◽  
Electrophoretic Mobilities ◽  
Chain Composition ◽  
Normal Human

SummaryThe polypeptide chain composition of fibrinogen and cross- linked fibrin from normal platelets and plasma has been compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Fibrinogen was prepared from a lysate of normal human platelets by ethanol precipitation and by antifibrinogen immunoaffinity chromatography. While the Aα chains of platelet fibrinogen appeared to be somewhat degraded, the electrophoretic mobilities of purified platelet and plasma fibrinogen Bβ an γ chains were the same. Crosslinked fibrin was prepared from platelet and plasma fibrinogen and γ chain dimers identified by their electrophoretic mobility, incorporation of the lysine analog dansyl cadaverine during crosslinking, and by reaction with antifibrinogen antibody in Western blots. Platelet crosslinked fibrin had a different polypeptide chain composition than that of plasma crosslinked fibrin with absence of the γ50–γ57.5 dimer in the platelet fibrin. This finding indicates that the γ57.5 chain, which constitutes 5% of total normal plasma fibrinogen γ chains, is either absent or present in markedly reduced amounts in platelet fibrinogen.

scholarly journals Lectin-Like Glycoprotein PsNLEC-1 Is Not Correctly Glycosylated and Targeted in Boron-Deficient Pea Nodules

2001 ◽  
Vol 14 (5) ◽  
pp. 663-670 ◽  
Author(s):  
Luis Bolaños ◽  
Arancha Cebrián ◽  
Miguel Redondo-Nieto ◽  
Rafael Rivilla ◽  
Ildefonso Bonilla
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Root Nodules ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Host Cells ◽  
Immunogold Localization ◽  
Electrophoretic Mobilities ◽  
Cytoplasmic Vesicles ◽  
Infected Cells

Symbiosome development was studied in pea root nodules from plants growing in the absence of boron (B). Rhizobia released into the host cells of nodules from B-deficient plants developed to abnormal endophytic forms with an altered electrophoretic lipopolysaccharide pattern. Immunostaining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotting of nodule homogenates with antibodies that recognize glycoprotein components showed that two previously described lectin-like glycoproteins (PsNLEC-1A and PsNLEC-1B) did not harbor the carbohydrate epitope normally recognized by specific monoclonal antibodies. Material derived from B-deficient nodules, however, still contained three antigenic isoforms with similar electrophoretic mobilities to PsNLEC-1 isoforms A, B, and C. These could be detected following immunoblotting and immunostaining with a specific antiserum originating from the purified PsNLEC protein that had been heterologously expressed in Escherichia coli. Immunogold localization of PsNLEC-1 sugar epitopes in B-deficient nodules showed that they were associated mostly with cytoplasmic vesicles rather than normal localization in the symbiosome compartment of mature infected cells. These results suggest that a modification of the glycosyl-moieties of PsNLEC-1 and an alteration of vesicle targeting occur during the development of pea nodules in the absence of B, and that these changes are associated with the development of aberrant nonfunctional symbiosomes.

scholarly journals Arginase from human full-term placenta

1976 ◽  
Vol 159 (3) ◽  
pp. 579-583 ◽  
Author(s):  
R Porta ◽  
C Esposito ◽  
A Martin ◽  
G D Pietra
Keyword(s):  
Gel Electrophoresis ◽  
Competitive Inhibition ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Disc Electrophoresis ◽  
Full Term ◽  
Term Placenta ◽  
A Value ◽  
Optimum Ph

Arginase was purified about 1800-fold from extracts of human full-term placenta; the enzyme appeared to be homogenous by disc electrophoresis and molecular-sieve chromatography. The mol. wt. determination by gel filtration and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis yielded a value of 70000 for the most pure and the partially purified enzyme. The human placenta arginase is a metalloenzyme with an optimum pH of 9.1. The Km for L-arginine is 27 mM. L-Ornithine and L-lysine show competitive inhibition with Ki values of 6.3 and 14 mM respectively.

scholarly journals Protein Profile Study of Some Nigerian Sesame (Sesamum indicum L.) Accessions

2015 ◽  
Vol 3 (2) ◽  
pp. 322-329 ◽  
Author(s):  
Gbenga Olorunshola Alege
Keyword(s):  
Genetic Diversity ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sesamum Indicum ◽  
Genomic Changes ◽  
Sds Page ◽  
Sesame Genotypes ◽  
Species Specific ◽  
Total Seed

This study was carried out to investigate the genetic diversity among 23 sesame (Sesamum indicum L.) accessions obtained from different agro-ecological localities from 10 different states across 4 geopolitical zones in Nigeria using evidence from Sodium Dodecyl Polyacrylamide Gel Electrophoresis (SDS-PAGE). Total seed protein of the studied plants resolved on 12% SDS-PAGE showed variations in numbers and intensity of bands among the different sesame accessions. Thirteen (13) major bands were recorded in this study. Lack of unique band and presence of common band (band 7) among the 23 studied sesame accessions indicate some levels of genetic affinity and evidence of common evolutionary origin of the sesame genotypes. This band can therefore be tagged as species specific band for discriminating Sesamum indicum. Cluster analysis grouped the 23 sesame genotypes into two clusters with similarity coefficient ranging from 0.42 to 0.96 which indicates existence of genetic diversity; therefore there is ample opportunity for improving the 23 sesame genotypes. Variations in protein bands observed among the 23 studied plants could be attributed to genomic changes taken place during species diversification. It can be concluded that genetic diversity existed among Nigerian sesame for the improvement of characters of interest. Accessions 9 (YOL), 15(OTT), 22 (OFF) and 23 (JAL) are therefore recommended for used in future breeding programs for the development of improved sesame varieties.Int J Appl Sci Biotechnol, Vol 3(2): 322-329 DOI: http://dx.doi.org/10.3126/ijasbt.v3i2.12734

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