Purification of chromaffin granule membrane glycoproteins by affinity chromatography on lectin columns

1985 ◽  
Vol 13 (4) ◽  
pp. 710-711 ◽  
Author(s):  
SARAH L. WOOD ◽  
DAVID K. APPS ◽  
JOHN H. PHILLIPS
Keyword(s):  
Affinity Chromatography ◽  
Membrane Glycoproteins ◽  
Chromaffin Granule ◽  
Granule Membrane

scholarly journals Fractionation of membrane proteins by temperature-induced phase separation in Triton X-114. Application to subcellular fractions of the adrenal medulla

1986 ◽  
Vol 233 (2) ◽  
pp. 525-533 ◽  
Author(s):  
J G Pryde ◽  
J H Phillips
Keyword(s):  
Endoplasmic Reticulum ◽  
Phase Separation ◽  
Membrane Proteins ◽  
Low Temperature ◽  
Membrane Glycoproteins ◽  
Chromaffin Granule ◽  
Rich Phase ◽  
Granule Membrane ◽  
Separation Of Proteins ◽  
Triton X

After solubilization with the detergent Triton X-114, membrane proteins may be separated into three groups: if the membrane is sufficiently lipid-rich, one family of hydrophobic constituents separates spontaneously at low temperature; warming at 30 degrees C leads to separation of a detergent-rich phase and an aqueous phase. Using the chromaffin-granule membrane as a model, we found that many intrinsic membrane glycoproteins are found in the latter phase, probably maintained in solution by adherent detergent. They precipitate, however, when this is removed by dialysis, leaving in solution those truly hydrophilic proteins that were originally adhering to the membranes. We have used this method with mitochondria, and with Golgi- and rough-endoplasmic-reticulum-enriched microsomal fractions: it has proved to be a rapid and convenient method for effecting a partial separation of proteins from a variety of different membranes.

Poly(N-Acetyllactosaminyl) Oligosaccharides of Chromaffin Granule Membrane Glycoproteins

1988 ◽  
Vol 51 (6) ◽  
pp. 1819-1824 ◽  
Author(s):  
Richard U. Margolis ◽  
Reiner Fischer-Colbrie ◽  
Renée K. Margolis
Keyword(s):  
Membrane Glycoproteins ◽  
Chromaffin Granule ◽  
Granule Membrane

scholarly journals Determination of the proportion of sealed vesicles in a preparation of chromaffin granule membrane 'ghosts'

FEBS Letters ◽  
1982 ◽  
Vol 144 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Anne Hunter ◽  
Keith Waldron ◽  
David K. Apps
Keyword(s):  
Chromaffin Granule ◽  
Granule Membrane

Antibodies Against Platelet Membrane Glycoproteins: Effect On Ristocetin-Induced Platelet Aggregation

1981 ◽  
Author(s):  
E F Ali-Briggs ◽  
C S P Jenkins ◽  
K J Clemetson
Keyword(s):  
Platelet Aggregation ◽  
Affinity Chromatography ◽  
Platelet Membrane ◽  
Receptor Activity ◽  
Free Medium ◽  
Membrane Glycoproteins ◽  
Lectin Affinity Chromatography ◽  
Lectin Affinity ◽  
Fab Fragments ◽  
Induced Aggregation

Some membrane glycoproteins (GPs) have been isolated by lectin-affinity chromatography and antibodies towards them have been raised. Platelets that have lost glycocalicin no longer respond to ristocetin-human VIII:WF, bovine VIIIR:WF, or to anti-glycocalicin or anti-GPs la and lb antibodies but are still agglutinated by anti-GPs lib and Ilia antibodies. Anti-GPs la and lb and anti-glycocalicin antibodies, IgG and Fab' fragments inhibited ristocetin- human VIIIR:WF- and bovine VIIIR:WF-induced aggregation of fixed, washed platelets and of platelets in plasma while anti-GPs Hb and Ilia antibodies were without effect.Crossed immunoelectrophorectic studies showed that glycocalicin was present on whole platelets in only trace amounts; anti-glycocalicin antibodies, however, recognized a slower migrating component. Platelets incubated in an EDTA-free medium no longer respond to ristocetin-human VIIIRrWF. Membranes isolated from such platelets contained glycocalicin which cross-reacted with a remnant of the slower migrating component. Anti-GPs la and lb antibodies gave more complex patterns but it was possible to identify the slower moving component recognized by the anti-glycocalicin antibodies.These results show that glycocalicin is not normally found as such on whole platelets but is present as a precursor which is most likely GP lb. On degradation of this precursor, glycocalicin is released from the membrane and VIIIRrWF-receptor activity is lost.

scholarly journals PADGEM protein in human erythroleukemia cells

Blood ◽  
1989 ◽  
Vol 73 (3) ◽  
pp. 722-728 ◽  
Author(s):  
E Yeo ◽  
BC Furie ◽  
B Furie
Keyword(s):  
Molecular Weight ◽  
Binding Sites ◽  
Protein A ◽  
Flow Cytometric Analysis ◽  
Surface Expression ◽  
Membrane Glycoproteins ◽  
Surface Binding ◽  
Cell Content ◽  
Granule Membrane ◽  
Hel Cells

Abstract PADGEM protein, a platelet alpha granule membrane glycoprotein with a molecular weight of 140,000, is translocated to the plasma membrane during granule secretion and platelet activation. PADGEM protein is expressed on the surface of activated platelets but not on the surface of resting platelets. Human erythroleukemia (HEL) cells contain platelet alpha granule-like organelles, alpha granule proteins, and express platelet membrane glycoproteins GPIIb/IIIa and GPIb. We demonstrate that HEL cells express a protein that has a molecular weight identical to that of PADGEM and binds to anti-PADGEM antibodies. The exposure of HEL cells in culture to dimethylsulfoxide (DMSO) increased the number of cells expressing PADGEM. Fluorescence activated flow cytometric analysis demonstrated an increase in mean surface expression of PADGEM in DMSO-exposed cells compared to noninduced cells. Total cell content of PADGEM was increased 5.3-fold after DMSO exposure, as determined by radioimmunoassay. Direct binding experiments with the monoclonal anti-PADGEM antibody KC4 demonstrated specific, saturable, and time-dependent interaction of KC4 with HEL cells. A Kd of 7 nM was estimated. There were 14,000 surface binding sites per cell in noninduced cells and 24,000 surface binding sites per cell in DMSO- induced HEL cells. Surface expression of PADGEM protein on HEL cells was not increased with platelet agonists, including thrombin, epinephrine, ADP, nor cytokines, including IL-1, IL-2, tissue necrosis factor. The presence of PADGEM protein in HEL cells should facilitate the elucidation of the function of PADGEM protein.

Ganglioside Composition of Chromaffin Granule Membrane in Bovine Adrenal Medulla 1

1984 ◽  
Vol 95 (1) ◽  
pp. 155-160 ◽  
Author(s):  
Michiko SEKINE ◽  
Toshio ARIGA ◽  
Tadashi MIYATAKE ◽  
Yoichiro KURODA ◽  
Akemi SUZUKI ◽  
...  
Keyword(s):  
Adrenal Medulla ◽  
Chromaffin Granule ◽  
Bovine Adrenal Medulla ◽  
Granule Membrane ◽  
Ganglioside Composition

Reconstitution of the Mg2+ -ATPase of the chromaffin granule membrane

FEBS Letters ◽  
1979 ◽  
Vol 103 (2) ◽  
pp. 323-327 ◽  
Author(s):  
R.Martin Buckland ◽  
George K. Radda ◽  
Lalage M. Wakefield
Keyword(s):  
Chromaffin Granule ◽  
Granule Membrane
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