The similarity of histones from turtle erythrocytes and liver

1981 ◽  
Vol 59 (4) ◽  
pp. 273-279 ◽  
Author(s):  
R. G. Rutledge ◽  
C. E. Shay ◽  
G. L. Brown ◽  
J. M. Neelin
Keyword(s):  
Gel Electrophoresis ◽  
Minor Component ◽  
Sea Turtle ◽  
Exchange Chromatography ◽  
The Other ◽  
Cation Exchange Chromatography ◽  
Mammalian Tissues ◽  
Exclusion Chromatography ◽  
A Minor ◽  
Molecular Exclusion Chromatography

Using the fresh-water "red-eared turtle" Pseudomys scriptans elegans, we have confirmed the existence of a minor component H1s among the lysine-rich histones of turtle erythrocytes by three forms of gel electrophoresis and two forms of chromatography. It was separated from the major components by both cation-exchange chromatography and molecular-exclusion chromatography and shown to differ slightly but significantly in content of several amino acids as well as being shorter than the other lysine-rich histones. Although its composition is close to that of the "tissue-specific" F1b of sea turtle erythrocytes, it is present in even greater relative amount in red-eared turtle livers. In most respects it resembles the satellite histone H10 of mammalian tissues. No unusual histones were observed among the perchloric acid insoluble histones of turtle erythrocytes.

The Kidney and Fibrinolysis

1970 ◽  
Vol 24 (01/02) ◽  
pp. 026-032 ◽  
Author(s):  
N. A Marsh
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Enzyme System ◽  
Normal Animal ◽  
Fibrinolytic Enzyme ◽  
The Other ◽  
Exclusion Chromatography ◽  
Normal Urine ◽  
Renal Origin ◽  
Molecular Exclusion Chromatography

SummaryMolecular exclusion chromatography was performed on samples of urine from normal and aminonucleoside nephrotic rats. Normal urine contained 2 peaks of urokinase activity, one having a molecular weight of 22,000 and the other around 200,000. Nephrotic urine contained three peaks of activity with MW’s 126,000, 60,000 and 30,000. Plasma activator determined from euglobulin precipitate had a MW. in excess of 200,000. The results indicate that in the normal animal, plasma plasminogen activator does not escape into the urine in substantial quantities but under the conditions of extreme proteinuria there may be some loss through the kidney. The alteration in urokinase output in nephrotic animals indicates a greatly disordered renal fibrinolytic enzyme system.The findings of this study largely support the hypothesis that plasma plasminogen activator of renal origin and urinary plasminogen activator (urokinase) are different molecular species.

A modified fractionation procedure for avian erythrocyte histones

1969 ◽  
Vol 47 (12) ◽  
pp. 1121-1123 ◽  
Author(s):  
G. H. M. Adams ◽  
J. M. Neelin
Keyword(s):  
Cation Exchange ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Fractionation Procedure ◽  
Exclusion Chromatography ◽  
One Step ◽  
Avian Erythrocyte ◽  
Fractionation Method ◽  
Time Required

The fractionation method of Vidali and Neelin for avian erythrocyte histones was modified to reduce the time required to obtain clean fractions of serine-rich and arginine-rich histones. Histones were extracted from washed nuclei in one step with 0.20 M HCl. The lysine-rich and the moderately lysine-rich histones were fractionated by cation-exchange chromatography but the serine-rich and arginine-rich histones were eluted together. These histones were separated by subsequent exclusion chromatography.

scholarly journals Isolation and physical characterization of the MUC7 (MG2) mucin from saliva: evidence for self-association

1998 ◽  
Vol 334 (2) ◽  
pp. 415-422 ◽  
Author(s):  
Ravi MEHROTRA ◽  
David J. THORNTON ◽  
John K. SHEEHAN
Keyword(s):  
Molecular Mass ◽  
Peptide Mapping ◽  
Minor Component ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Gel Chromatography ◽  
Guanidinium Chloride ◽  
Average Molecular Mass ◽  
Self Association ◽  
A Minor

Saliva contains two major families of mucins (MG1 and MG2); the polypeptide of the smaller of these glycoproteins (MG2) has been assigned as the product of the MUC7 gene. In this study we have devised a rapid two-step procedure that recovers this glycoprotein essentially free of other components and in sufficient quantity to enable physical and self-interaction studies. Raw saliva was solubilized in 4 M guanidinium chloride and thereafter subjected to Sepharose CL-4B chromatography. The MG2-rich fraction was recovered free from the larger MG1 glycoproteins and also smaller proteins/glycoproteins (molecular mass less than 100 kDa). MG2 glycoproteins were finally purified by anion-exchange chromatography on Mono Q. The purity of the preparation was assessed by SDS/PAGE after radiolabelling of the molecules with [14C]acetic anhydride. Peptide mapping, N-terminal sequencing and amino acid analysis verified the polypeptide of the mucins as the MUC7 gene product. The isolated molecules were examined by electron microscopy and appeared as short flexible worm-like structures 30–120 nm in length. The distribution was heterogeneous, containing a major component with number-average and weight-average lengths of 52 and 55 nm respectively and a minor component with number-average and weight-average lengths of 94 and 98 nm respectively. We propose that the two differently sized populations represent monomeric and dimeric species of the mucins. Gel chromatography performed in 0.2 M NaCl indicated the presence of monomers, dimers and tetramers; an average molecular mass for the preparation was 192 kDa. However, in 4 M guanidinium chloride the molecular mass was 158 kDa and a similar molecular mass (155 kDa) was determined for the mucin preparation after reduction. These results suggest that the mucins might self-associate via a protein-mediated interaction. On the basis of the results a model is proposed for the self-association of the MUC7 mucin, which might be important for its biological function.

scholarly journals Murine B-cell subpopulations responsive to T-dependent and T-independent antigens.

1976 ◽  
Vol 144 (2) ◽  
pp. 382-397 ◽  
Author(s):  
G K Lewis ◽  
R Ranken ◽  
D E Nitecki ◽  
J W Goodman
Keyword(s):  
B Cells ◽  
B Cell ◽  
Minor Component ◽  
Memory Cell ◽  
Keyhole Limpet Hemocyanin ◽  
The Other ◽  
Other Hand ◽  
Cell Mitogen ◽  
A Minor

Strain A/J mice made secondary indirect plaque-forming cell (PFC) responses to azobenzenearsonate (ABA) conjugates of giant keyhole limpet hemocyanin (KLH), a thymic-dependent antigen, but not to conjugates of Ficoll, a T-independent antigen. ABA-Ficoll was also unable to elicit a response in animals primed with ABA-KLH, which have an expanded anti-ABA memory cell pool. On the other hand, ABA-Ficoll rendered mice unresponsive to ABA-KLH when administered before priming or boosting with the T-dependent immunogen. Hence, the T-independent antigen was able to tolerize but unable to trigger B-memory cells responsive to the T-dependent antigen. A/J mice immunized with dinitrophenyl conjugates of Ficoll or bovine IgG (BGG) made vigorous IgM and IgG PFC responses. PFC responses to ABA-KLH and 2,4-dinitrophenyl (DNP)-BGG were abrogated by depleting mice of C3 with cobra venom factor, whereas the IgM and IgG PFC responses to DNP-Ficoll were unaffected. B lymphocytes were fractionated on the basis of receptors for C3 and the subpopulations were assayed for in vitro PFC responses to DNP-Ficoll. Very little response was obtained from complement receptor lymphocyte [CRL(+)] B cells, whereas CRL(-) cells were more responsive than unfractionated B cells. Both populations responded to a polyclonal B-cell mitogen (lipopolysaccharide). On the other hand, the in vitro PFC response to a T-dependent antigen (sheep erythrocytes) correlated with the presence of CRL(+) B cells in the cultures. However, a minor component of this response, sensitive to anti-Thy-1 serum, was made by CRL(-) B cells, indicating the existence of subpopulations of T-dependent B cells with different signalling requirements. The results suggest that most B cells responsive to T-dependent antigens possess receptors for C3 and that C3 plays an obligatory role in the response of these cells. A distinct subpopulation of B cells which lack C3 receptors respond to T-independent antigens. The precursors of PFC for the ABA epitope reside largely or exclusively in the CRL(+) compartment in A/J mice, whereas precursors for the DNP determinant are found in both compartments.

scholarly journals Low-Mr iron isolated from guinea pig reticulocytes as AMP-Fe and ATP-Fe complexes

1989 ◽  
Vol 261 (3) ◽  
pp. 787-792 ◽  
Author(s):  
J Weaver ◽  
S Pollack
Keyword(s):  
Guinea Pig ◽  
Anion Exchange ◽  
Dominant Species ◽  
Exchange Resin ◽  
Anion Exchange Resin ◽  
Minor Component ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Fe Complexes ◽  
A Minor

Guinea pig reticulocytes were pulse-labelled with 59Fe bound to transferrin. Haemolysates prepared from these reticulocytes were subjected to rapid (NH1)2SO1 precipitation and then chromatography on an anion-exchange resin. ATP-bound 59Fe was the dominant species in the reticulocyte cytosol; 2,3-bisphosphoglycerate and GTP iron complexes were not detected despite the fact that these were stable with (NH1)2SO1 precipitation and readily detected with anion-exchange chromatography. AMP-bound Fe was a minor component of the cytosol following rapid (NH1)2SO4 precipitation, and the major component when iron was released from transferrin by haemolysates. We speculate that ATP-Fe may be degraded in the cell to permit utilization of its iron for haem synthesis.

Comparison of the histones from fish erythrocytes

1977 ◽  
Vol 55 (12) ◽  
pp. 1220-1227 ◽  
Author(s):  
Brian L. A. Miki ◽  
James M. Neelin
Keyword(s):  
Cation Exchange ◽  
Polyacrylamide Gel Electrophoresis ◽  
Exchange Chromatography ◽  
White Sucker ◽  
Cation Exchange Chromatography ◽  
Apparent Absence ◽  
Histone Fraction ◽  
Functional Homology ◽  
Diverse Species ◽  
Exclusion Chromatography

Because of contentions concerning the generality of cell-specific histone 5 (H5) in nucleated erythrocytes of species other than birds, the erythrocyte histones of carp, trout, perch, black crappie, and white sucker were studied under conditions which minimize proteolysis. All were examined by polyacrylamide gel electrophoresis, and histone 1 (H1) and H5 were purified by cation-exchange chromatography on Amberlite CG-50 and by exclusion chromatography through BioGel P60.A protein homologous to H5 could be isolated and identified from the mature erythrocytes of carp, trout, perch, and black crappie but not from white sucker. The relative amounts of H5 differed extensively and inversely in proportion to H1, implying some functional homology between these proteins. The extremely variable levels of H5, including its apparent absence from one species, suggests that typical H5 is not essential to the function or development of nucleated erythrocytes.Although H1 is the most divergent histone fraction, H5 is also highly variable. Fish erythrocyte H5 differs from avian H5 in relative electrophoretic mobility, ease of elution from Amberlite CG-50 cation-exchange columns, and amino acid composition. H5's from fish tend to have more threonine but less serine, arginine, glutamic acid, and histidine than avian H5's. Fish H5's are more diverse than avian H5's and resemble the H5 homologues from other extremely diverse species; thus avian H5 may be an extreme in specialization of an H1 subfraction.

Interferences appearing in fluorometrically measured liquid-chromatographic profiles of creatine kinase isoenzymes in serum.

1980 ◽  
Vol 26 (6) ◽  
pp. 707-711 ◽  
Author(s):  
T D Schlabach ◽  
J A Fulton ◽  
P B Mockridge ◽  
E C Toren
Keyword(s):  
Creatine Kinase ◽  
High Performance ◽  
Minor Component ◽  
The Other ◽  
Present Evidence ◽  
Human Sera ◽  
A Minor ◽  
Liquid Chromatographic ◽  
Measured Liquid ◽  
Chromatographic Profiles

Abstract We observed nonenzymic peaks when serum isoenzymes of lactate dehydrogenase (EC 1.1.1.27; LD) and creatine kinase (EC 2.7.3.2.; CK) were separated by “high-performance” liquid chromatography and detected by continuously monitoring the column effluent for enzyme activity. Such background peaks were particularly apparent in CK isoenzyme profiles obtained from human sera. We observed two nonenzymic peaks with fluorescence detection, one in the CK-MB region, the other in the CK-BB region. Serum albumin was a major component in the artifactual CK-MB peak, with lipoprotein as a minor component. We present evidence that the material responsible for the other peak fluoresced quite strongly and is mostly pre-albumin.

scholarly journals Two subsets of human T lymphocytes expressing gamma/delta antigen receptor are identifiable by monoclonal antibodies directed to two distinct molecular forms of the receptor.

1988 ◽  
Vol 168 (2) ◽  
pp. 491-505 ◽  
Author(s):  
C Bottino ◽  
G Tambussi ◽  
S Ferrini ◽  
E Ciccone ◽  
P Varese ◽  
...  
Keyword(s):  
Target Cell ◽  
Peripheral Blood ◽  
Minor Component ◽  
The Other ◽  
Molecular Forms ◽  
Gamma Delta ◽  
Gamma Receptor ◽  
Other Hand ◽  
Reducing Conditions ◽  
A Minor

Two mAbs directed to the TCR-gamma/delta were analyzed for their pattern of reactivity with CD3+WT31- cell populations or clones. In normal individuals, the BB3 mAb reacted with approximately 2/3 of peripheral blood CD3+WT31- lymphocytes, whereas delta-TCS-1 stained approximately 1/3 of such cells. In addition, the sum of the percentages of BB3+ and delta-TCS-1+ cells approximated the percentages of peripheral blood CD3+WT31- lymphocytes in seven normal donors tested. Also, in peripheral blood-derived polyclonal CD3+WT31- populations, cultured in IL-2, cells reacting with one or another mAb accounted for the whole cell population. On the other hand, only delta-TCS-1-reactive cells, but not BB3+ cells, could be detected in unfractionated as well as in CD4-8-thymocyte populations. Analysis of peripheral blood-derived CD3+WT31- clones showed that 70% of 72 clones analyzed reacted with BB3 mAb, but not with delta-TCS-1 mAb. On the other hand, delta-TCS-1 mAb stained the remaining BB3- clones. Five clones expressing medium-low amounts of CD8 antigen were BB3- delta-TCS-1+. Both types of clones lysed the Fc gamma receptor-bearing P815 target cell in the presence of anti-CD3 mAb (but not of mAb directed against HLA-DR, CD7 molecules, or TCR-alpha/beta). In this cytolytic assay, BB3 mAb induced target cell lysis only by BB3+ clones, whereas delta-TCS-1 mAb was effective only with delta-TCS-1+ clones. The CD3-associated surface molecules expressed by BB3+ or delta-TCS-1+ clones were analyzed after cell surface iodination and immunoprecipitation with the corresponding anti-TCR mAb or with anti-CD3 mAb (in digitonin-containing buffer). In SDS-PAGE, molecules immunoprecipitated from 13 BB3+ clones displayed, under nonreducing conditions, a molecular weight of 80 kD (in some cases, a minor 38-kD band could be detected). Under reducing conditions, two major components of 44 and 41 kD (and a minor component of 38 kD) were detected. On the other hand, TCR molecules immunoprecipitated from 11 different delta-TCS-1+ clones appeared as a diffuse band of 41-44 kD, both under reducing and nonreducing conditions (under non-reducing condition, an additional 38-kD band was present). Therefore, BB3+ cells express a disulphide-linked form of TCR-gamma/delta whereas delta-TCS-1+ cells express a non-disulphide-linked form.(ABSTRACT TRUNCATED AT 400 WORDS)

Three assays for glycohemoglobin compared

1995 ◽  
Vol 41 (2) ◽  
pp. 191-195 ◽  
Author(s):  
U Turpeinen ◽  
U Karjalainen ◽  
U H Stenman
Keyword(s):  
Cation Exchange ◽  
Correlation Coefficient ◽  
Reference Method ◽  
Hplc Method ◽  
Exchange Chromatography ◽  
The Other ◽  
Affinity Binding ◽  
Cation Exchange Chromatography ◽  
The Difference

Abstract Using 123 specimens, we compared the concordance of three different methods for determining glycohemoglobin (GHb): the Diamat (Bio-Rad Laboratories), an automated analyzer measuring HbA1c by cation-exchange chromatography; an assay with the IMx analyzer (Abbott Laboratories), based on boronate affinity binding; and an HPLC method measuring HbA1c by cation-exchange chromatography on a PolyCAT A column (PolyLC Inc.). The Pearson's correlation coefficient between PolyCAT A and Diamat was 0.900 +/- 0.038 (mean +/- 2 SD) and between PolyCAT A and IMx, 0.857 +/- 0.042. However, up to twofold differences were seen in some samples. The proportion of GHb was consistently lower with the PolyCAT A method than with the other two assays, apparently because of better separation of HbA1c from nonglycated coeluting forms of Hb. The difference in glycation percentage between the PolyCAT A and Diamat methods is 2-3% over the whole concentration range. These results point to the limitations of Diamat as a reference method to be used to calibrate other methods for determining HbA1c. Further, a switch from one method to another is likely to cause considerable problems in the clinical follow-up of certain patients.

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