scholarly journals Identification of protein binding sites in genomic DNA by two-dimensional gel electrophoresis

1991 ◽  
Vol 19 (7) ◽  
pp. 1369-1374 ◽  
Author(s):  
Andrea Boffini ◽  
Pierre Prentki
Keyword(s):  
Protein Binding ◽  
Binding Sites ◽  
Gel Electrophoresis ◽  
Genomic Dna ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Protein Binding Sites

Two-dimensional gel selection of protein binding sites on DNA

1993 ◽  
Vol 14 (1) ◽  
pp. 259-265 ◽  
Author(s):  
Andrea Boffini ◽  
Pierre Prentki
Keyword(s):  
Protein Binding ◽  
Binding Sites ◽  
Two Dimensional ◽  
Protein Binding Sites ◽  
Selection Of

scholarly journals Analysis of Bacterial Populations in the Environment Using Two-dimensional Gel Electrophoresis of Genomic DNA and Complementary DNA

2011 ◽  
Vol 26 (2) ◽  
pp. 184-187 ◽  
Author(s):  
Guo-Hua Liu ◽  
Tatsuo Nakamura ◽  
Takashi Amemiya ◽  
Narasimmalu Rajendran ◽  
Kiminori Itoh
Keyword(s):  
Gel Electrophoresis ◽  
Genomic Dna ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Bacterial Populations ◽  
Complementary Dna

scholarly journals A platelet membrane glycoprotein (GP) deficiency in healthy blood donors: Naka- platelets lack detectable GPIV (CD36)

Blood ◽  
1990 ◽  
Vol 76 (9) ◽  
pp. 1698-1703 ◽  
Author(s):  
N Yamamoto ◽  
H Ikeda ◽  
NN Tandon ◽  
J Herman ◽  
Y Tomiyama ◽  
...  
Keyword(s):  
Binding Sites ◽  
Gel Electrophoresis ◽  
Blood Donors ◽  
Platelet Membrane ◽  
Platelet Glycoprotein ◽  
Membrane Glycoprotein ◽  
Two Dimensional ◽  
Platelet Concentrates ◽  
Two Dimensional Gel Electrophoresis ◽  
Crossed Immunoelectrophoresis

It has recently been shown that the Naka antigen, which is absent in 3% to 11% of Japanese blood donors, is expressed on platelet glycoprotein IV (GPIV; CD36) (Tomiyama et al, BLOOD, 75:684, 1990). In the present studies, flow cytometry was used to distinguish differences in the reactivity of Naka+ and Naka- platelets with both OKM5, a monoclonal antibody that recognizes an epitope on GPIV, and with polyclonal anti- GPIV antibody. OKM5 was also used to screen 871 platelet concentrates prepared from healthy US blood donors. Three of these showed markedly deficient binding of 125I-OKM5 or an incidence of 0.34%. Two of these donors were re-accessed and showed less than 1% binding of 125I-OKM5 as compared with 10,300 +/- 1,500 binding sites per platelet in controls (n = 4). Platelets from these two US donors were radiolabeled (125I, 3H) and compared with control platelets and with platelets from Japanese Naka+ and Naka- donors by crossed immunoelectrophoresis, protein blots, immunoprecipitation, and two-dimensional gel electrophoresis. GPIV could not be detected by any of these techniques in the Naka- platelets nor in the donors whose platelets showed deficient binding of OKM5. These results suggest that GPIV functions as an isoantigen rather than an alloantigen in immunizing Naka- platelet recipients. This is the first report of the absence of a major platelet membrane GP in healthy blood donors.

scholarly journals A platelet membrane glycoprotein (GP) deficiency in healthy blood donors: Naka- platelets lack detectable GPIV (CD36)

Blood ◽  
1990 ◽  
Vol 76 (9) ◽  
pp. 1698-1703 ◽  
Author(s):  
N Yamamoto ◽  
H Ikeda ◽  
NN Tandon ◽  
J Herman ◽  
Y Tomiyama ◽  
...  
Keyword(s):  
Binding Sites ◽  
Gel Electrophoresis ◽  
Blood Donors ◽  
Platelet Membrane ◽  
Platelet Glycoprotein ◽  
Membrane Glycoprotein ◽  
Two Dimensional ◽  
Platelet Concentrates ◽  
Two Dimensional Gel Electrophoresis ◽  
Crossed Immunoelectrophoresis

Abstract It has recently been shown that the Naka antigen, which is absent in 3% to 11% of Japanese blood donors, is expressed on platelet glycoprotein IV (GPIV; CD36) (Tomiyama et al, BLOOD, 75:684, 1990). In the present studies, flow cytometry was used to distinguish differences in the reactivity of Naka+ and Naka- platelets with both OKM5, a monoclonal antibody that recognizes an epitope on GPIV, and with polyclonal anti- GPIV antibody. OKM5 was also used to screen 871 platelet concentrates prepared from healthy US blood donors. Three of these showed markedly deficient binding of 125I-OKM5 or an incidence of 0.34%. Two of these donors were re-accessed and showed less than 1% binding of 125I-OKM5 as compared with 10,300 +/- 1,500 binding sites per platelet in controls (n = 4). Platelets from these two US donors were radiolabeled (125I, 3H) and compared with control platelets and with platelets from Japanese Naka+ and Naka- donors by crossed immunoelectrophoresis, protein blots, immunoprecipitation, and two-dimensional gel electrophoresis. GPIV could not be detected by any of these techniques in the Naka- platelets nor in the donors whose platelets showed deficient binding of OKM5. These results suggest that GPIV functions as an isoantigen rather than an alloantigen in immunizing Naka- platelet recipients. This is the first report of the absence of a major platelet membrane GP in healthy blood donors.

Diversity of Glycoprotein Deficiencies in Glanzmann’s Thrombasthenia

1985 ◽  
Vol 54 (03) ◽  
pp. 626-629 ◽  
Author(s):  
M Meyer ◽  
F H Herrmann
Keyword(s):  
High Resolution ◽  
Gel Electrophoresis ◽  
Type Ii ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Structural Variant ◽  
Glanzmann’S Thrombasthenia ◽  
Crossed Immunoelectrophoresis ◽  
Glanzmann's Thrombasthenia ◽  
Platelet Proteins

SummaryThe platelet proteins of 9 thrombasthenic patients from 7 families were analysed by high resolution two-dimensional gel electrophoresis (HR-2DE) and crossed immunoelectrophoresis (CIE). In 7 patients both glycoproteins (GPs) IIb and Ilia were absent or reduced to roughly the same extent. In two related patients only a trace of GP Ilb-IIIa complex was detected in CIE, but HR-2DE revealed a glycopeptide in the position of GP Ilia in an amount comparable to type II thrombasthenia. This GP Ilia-like component was neither recognized normally by anti-GP Ilb-IIIa antibodies nor labeled by surface iodination. In unreduced-reduced two-dimensional gel electrophoresis two components were observed in the region of GP Ilia. The assumption of a structural variant of GP Ilia in the two related patients is discussed.

EVALUATION OF TISSUE STEROID BINDING IN VITRO

1971 ◽  
Vol 68 (1_Suppl) ◽  
pp. S223-S246 ◽  
Author(s):  
C. R. Wira ◽  
H. Rochefort ◽  
E. E. Baulieu
Keyword(s):  
Protein Binding ◽  
Binding Sites ◽  
Experimental System ◽  
Specific Protein ◽  
Coupling Mechanism ◽  
Target Tissue ◽  
Protein Binding Sites ◽  
Definition Of ◽  
Hormone Specificity

ABSTRACT The definition of a RECEPTOR* in terms of a receptive site, an executive site and a coupling mechanism, is followed by a general consideration of four binding criteria, which include hormone specificity, tissue specificity, high affinity and saturation, essential for distinguishing between specific and nonspecific binding. Experimental approaches are proposed for choosing an experimental system (either organized or soluble) and detecting the presence of protein binding sites. Techniques are then presented for evaluating the specific protein binding sites (receptors) in terms of the four criteria. This is followed by a brief consideration of how receptors may be located in cells and characterized when extracted. Finally various examples of oestrogen, androgen, progestagen, glucocorticoid and mineralocorticoid binding to their respective target tissues are presented, to illustrate how researchers have identified specific corticoid and mineralocorticoid binding in their respective target tissue receptors.

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