scholarly journals Studies on the subcellular localization of human neutrophil alkaline phosphatase

1979 ◽  
Vol 36 (1) ◽  
pp. 401-412
Author(s):  
G.J. Rustin ◽  
P.D. Wilson ◽  
T.J. Peters
Keyword(s):  
Alkaline Phosphatase ◽  
Endoplasmic Reticulum ◽  
Density Gradient ◽  
Human Neutrophil ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Human Neutrophils ◽  
Gamma Glutamyl Transferase ◽  
Gamma Glutamyl ◽  
Glutamyl Transferase

The intracellular localization of alkaline phosphatase has been determined in human neutrophils with analytical subcellular fractionation by density gradient centrifugation and EM cytochemistry. Centrifugation on sucrose gradients containing 1 mM DETA and 5 units/ml of heparin showed that alkaline phosphatase was associated with a membranous component distinct from plasma membrane, mitochondria, specific granules and azurophil granules. There was no resolution from the endoplasmic reticulum. Density gradient centrifugation on a sucrose-imidazole-heparin gradient showed a clear resolution of the alkaline phosphatase-containing membranes from the Golgi and endoplasmic reticulum. Density gradient centrifugation of neutrophils that had been disrupted in the presenceof 0.12 mmol/l. digitonin clearly separated alkaline phosphatase-containing membranes from the endoplasmic reticulum. Part of the gamma-glutamyl transferase has a similar localization to that of alkaline phosphatase. EM cytochemistry of neutrophils, neutrophil homogenates and of the density gradient fractions identified alkaline phosphatase-containing granules as irregular-shaped, often tubular, structures. It is suggested that alkaline phosphatase and part of the gamma-glutamyl transferase activity are localized to a unique organelle in the human neutrophil.

scholarly journals Comparative Study of Serum Gamma Glutamyltransferase, 5’ Nucleotidase and Alkaline Phosphatase in Icteric and an-Icteric Biliary Disease Patients

2013 ◽  
Vol 12 (1) ◽  
Author(s):  
Dr. Maninder Pal Singh Gill
Keyword(s):  
Alkaline Phosphatase ◽  
Significant Rise ◽  
Control Group ◽  
P Value ◽  
Gamma Glutamyl Transferase ◽  
Biliary Disease ◽  
Gamma Glutamyl ◽  
Hepatobiliary Diseases ◽  
Glutamyl Transferase ◽  
Diagnostic Indicator

Introduction: Diagnostic enzymology plays a useful role in evaluation of various hepatobiliary diseases and numerous enzymes have been compared in different disorders. Among these, significance of Gamma Glutamyl Transferase and 5’ Nucleotidase over Alkaline Phosphatase has been stressed repeatedly, but mainly in the icteric obstructive biliary disease patients. In this study, these three enzymes were compared not only in the icteric but also the an-icteric biliary disease patients, particularly to look for elevation and significance of these enzymes in the latter group.Methods: The study was conducted on 50 biliary disease patients, who were further divided into an-icteric (32 patients) and icteric (18 patients) subgroups depending on their bilirubin levels. 50 subjects matched for age and sex with the study group were enrolled for the control group. Gamma Glutamyl Transferase, 5’Nucleotidase, Alkaline Phosphatase and bilirubin levels were evaluated in all the patients as well as the control subjects. Results: All three enzymes showed a significant rise in the icteric subgroup (p value < 0.001). However, in the an-icteric subgroup, only Gamma Glutamyl Transferase and 5’Nucleotidase showed a significant rise. The rise was more for Gamma Glutamyl Transferse (1.60 times normal, p < 0.001) as compared to 5’Nucleotidase (1.39 times normal, p < 0.01). Conclusion: Gamma Glutamyl Transferase and 5’Nucleotidase are useful for evaluation of not only obstructive biliary disease patients but also for the patients with biliary disease who are an-icteric, and out of these two, the former is a more valuable diagnostic indicator in such diseases

scholarly journals The locations of cathepsin activity and β-glucuronidase in the Guerin T8 tumour

1970 ◽  
Vol 118 (3) ◽  
pp. 543-549 ◽  
Author(s):  
A. R. Poole
Keyword(s):  
Endoplasmic Reticulum ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Differential Centrifugation ◽  
Unit Membrane ◽  
Electron Microscopic ◽  
Cathepsin Activity ◽  
Density Gradient Sedimentation

Tumour homogenate fractions, isolated by differential centrifugation, were subfractionated by density-gradient centrifugation. Biochemical and electron microscopic analyses revealed that β-glucuronidase and cathepsin activity were associated with a class (possibly two) of lysosomal particles of density greater than those of mitochondria and the endoplasmic reticulum. Lysosomes sedimented by low g forces were vacuolar, electron-dense, delineated by a unit membrane and about 0.2μm in diameter. β-Glucuronidase was also apparently associated with ribosomes whereas cathepsin was bound in part to the endoplasmic reticulum. Catalase and glucose 6-phosphatase possessed slightly different density-gradient sedimentation profiles.

scholarly journals Evidence for small intracellular vesicles in human blood phagocytes containing cytochrome b558 and the adhesion molecule CD11b/CD18

Blood ◽  
1993 ◽  
Vol 81 (11) ◽  
pp. 3122-3129 ◽  
Author(s):  
J Calafat ◽  
TW Kuijpers ◽  
H Janssen ◽  
N Borregaard ◽  
AJ Verhoeven ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Density Gradient ◽  
Whole Blood ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Surface Membrane ◽  
Ultrastructural Level ◽  
Human Neutrophils ◽  
Cytochrome B558 ◽  
Intracellular Vesicles

Human neutrophils contain a rapidly mobilizable pool of so-called secretory vesicles distinct from the azurophil granules and specific granules. Using human albumin as a marker for these intracellular vesicles in immuno-electron microscopy, we found that part of the cytochrome b558 in non-purified whole blood neutrophils colocalized in these vesicles. This was detected with monoclonal antibody (MoAb) CLB- 48, binding to the high molecular weight subunit of cytochrome b558. Approximately 65% of the albumin-containing vesicles showed MoAb CLB-48 labeling. This was also found in eosinophilic granulocytes and in monocytes. Cytofluorimetric determination of cytochrome b558 expression on the plasma membrane of intact, nonpurified granulocytes (and monocytes) with MoAb 7D5, which is directed against an extracellular epitope of cytochrome b558, did not show any binding. However, granulocytes (and monocytes) significantly bound 7D5 after density centrifugation. The positive binding of 7D5 to purified neutrophilic granulocytes correlated with a strongly reduced labeling of cytochrome b558 in the albumin-positive vesicles. Binding of CD11b MoAb CLB-B2.12 to the alpha subunit of the complement receptor type 3 (CR3) on the surface of intact, nonpurified neutrophils was detected to a limited extent in whole blood samples, but was strongly increased upon density gradient centrifugation of the cells, as we have described before. Investigation at the ultrastructural level showed that the CD11b antigen codistributed with albumin in vesicular structures in nonpurified phagocytes, especially in neutrophils and eosinophils. Together, these data substantiate the idea of an intracellular store that can be easily mobilized (even under the simple stress condition of density gradient centrifugation). Such mobilization may result in the expression of cytochrome b558 on the plasma membrane, as was indicated in this study. Apart from cytochrome b558, several other surface membrane molecules, as we show here for the integrin CD11b/CD18 (CR3), are probably also located in these rapidly mobilizable intracellular vesicles.

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