scholarly journals In resting conditions, the pancreatic granule membrane protein GP-2 is secreted by cleavage of its glycosylphosphatidylinositol anchor

1991 ◽  
Vol 277 (3) ◽  
pp. 879-881 ◽  
Author(s):  
E Paul ◽  
F A Leblond ◽  
D LeBel
Keyword(s):  
Membrane Protein ◽  
Phospholipase C ◽  
Exocrine Pancreas ◽  
Hydrophobic Properties ◽  
Glycosylphosphatidylinositol Anchor ◽  
Phosphodiester Bond ◽  
Granule Membrane ◽  
Anaesthetized Rats ◽  
Triton X ◽  
Hydrolysis Of

GP-2 is the major membrane protein of the exocrine pancreatic secretory granule. It is an integral protein which is anchored by a phosphatidylinositolglycan. In addition to being present in the soluble contents of the granule, GP-2 is also actively secreted by the pancreas. Although 93% of the GP-2 in the resting secretions of anaesthetized rats could be pelleted, Triton X-114 phase extraction showed that 70% of this GP-2 had lost its hydrophobic properties. Proteases have been postulated to release GP-2 from the membrane, but phospholipases also have the capacity to release the protein from the membrane by hydrolysis of its peculiar glycosylphosphatidylinositol membrane anchor. These studies show the presence of inositol 1,2-(cyclic)monophosphate on the secreted hydrophilic GP-2, confirming the involvement of an endogenous phospholipase C in the solubilization of GP-2 by the exocrine pancreas. It is therefore concluded that most of the GP-2 secreted by the pancreas of anaesthetized rats under resting conditions is released from the membrane by a phospholipase C which hydrolyses the phosphodiester bond linking GP-2 to its diradylglycerol anchor.

Influence of associated lipid on the properties of purified bovine erythrocyte acetylcholinesterase

1991 ◽  
Vol 69 (2-3) ◽  
pp. 154-162 ◽  
Author(s):  
Christine P. Nichol ◽  
Basil D. Roufogalis
Keyword(s):  
Phase Separation ◽  
Affinity Chromatography ◽  
Phospholipase C ◽  
Lectin Binding ◽  
Acetylcholinesterase Activity ◽  
Erythrocyte Membranes ◽  
Bovine Erythrocyte ◽  
Hydrophobic Properties ◽  
Native Form ◽  
Triton X

Acetylcholinesterase has been isolated from bovine erythrocyte membranes by affinity chromatography using a m-trimethylammonium ligand. The purified enzyme had hydrophobic properties by the criterion of phase partitioning into Triton X-114. The activity of the hydrophobic enzyme was seen as a slow-moving band in nondenaturing polyacrylamide gels. After treatment with phosphatidylinositol-specific phospholipase C, another form of active enzyme was produced that migrated more rapidly toward the anode in these gels. This form of the enzyme partitioned into the aqueous phase in Triton X-114 phase separation experiments and was therefore hydrophilic. The hydrophobic form bound to concanavalin A in the absence of Triton X-100. As this binding was partially prevented by detergent, but not by α-methyl mannoside, D-glucose, or myo-inositol, it is in part hydrophobic. Erythrocyte cell membranes showed acetylcholinesterase activity present as a major form, which was hydrophobic by Triton X-114 phase separation and in nondenaturing gel electrophoresis moved at the same rate as the purified enzyme. In the membrane the enzyme was more thermostable than when purified in detergent. The hydrophobic enzyme isolated, therefore, represents a native form of the acetylcholinesterase present in the bovine erythrocyte cell membrane, but in isolation its stability becomes dependent on amphiphile concentration. Its hydrophobic properties and lectin binding are attributable to the association with the protein of a lipid with the characteristics of a phosphatidylinositol.Key words: acetylcholinesterase, bovine erythrocytes, phosphatidylinositol-specific phospholipase C, phase separation, affinity chromatography.

scholarly journals Inhibition of glucose 6-phosphatase by pure and impure C-type phospholipases. Reactivation by phospholipid dispersions and protection by serum albumin

1975 ◽  
Vol 148 (2) ◽  
pp. 279-294 ◽  
Author(s):  
B R Cater ◽  
P Trivedi ◽  
T Hallinan
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Phospholipase C ◽  
Phosphatase Activity ◽  
Bovine Serum ◽  
Intact Control ◽  
Total Glucose ◽  
Triton X ◽  
Hydrolysis Of

1. Pure or impure C-type phospholipases hydrolysed rat liver microsomal phosphatides in situ at 5 degrees or 37 degrees C. At 5 degrees C mean hydrolysis of total phospholipids was 90% by Bacillus cereus and 75% by Clostridium perfringens (Clostridium welchii) C-type phospholipases. 2. Four degrees of inhibition of glucose 6-phosphatase (D-glucose 6-phosphate phosphohydrolase; EC 3.1.3.9) resulted. (a) At 37 degrees C inhibition was virtually complete and apparently irreversible. (b) At 5 degrees C phospholipase C inhibited 50-87% of the activity expressed by intact control microsomal fractions. (c) Bovine serum albumin present during delipidation alleviated most of this inhibition: at 5 degrees C phospholipase C plus bovine serum albumin inhibited by 0-35% (mean 18%):simultaneous stimulation by the destruction of its latency seems to offset glucose 6-phosphatase inhibition, sometimes completely. (d) If latency was first destroyed, phospholipase C plus bovine serum albumin inhibited 30-50% of total glucose 6-phosphatase activity at 5 degrees C. Only this inhibition is likely largely to reflect the lower availability of phospholipids, essential for maximal enzyme activity, as it is virtually completely reversed by added phospholipid dispersions. Co-dispersions of phosphatidylserine plus phosphatidylcholine (1:1, w/w) were especially effective but Triton X-100 was unable effectively to restore activity. 3. Considerable glucose 6-phosphatase activity survived 240min of treatment with phospholipase C at 5 degrees C, but in the absence of substrate or at physiological glucose 6-phosphate concentrations the delipidated enzyme was completely inactivated within 10min at 37 degrees C. However, 80mM-glucose 6-phosphate stabilized it and phospholipid dispersions substantially restored thermal stability. 4. It is concluded that glucose 6-phosphatase is at least partly phospholipid-dependent, and complete dependence is not excluded. For reasons discussed it is impossible yet to be certain which phospholipid class(es) the enzyme requires for activity.

scholarly journals Membrane Dipeptidase in the Pig Exocrine Pancreas: Ultrastructural Localization and Secretion

1998 ◽  
Vol 46 (7) ◽  
pp. 841-846 ◽  
Author(s):  
Denis LeBel ◽  
Gilles Grondin ◽  
Sarah Cook ◽  
Nigel M. Hooper
Keyword(s):  
Exocrine Pancreas ◽  
Ultrastructural Localization ◽  
Apical Plasma Membrane ◽  
Peptidase Activity ◽  
Apical Surface ◽  
Rich Phase ◽  
Granule Membrane ◽  
Membrane Bound ◽  
Triton X ◽  
Dipeptidase Activity

The GPI-anchored membrane dipeptidase is the major peptidase activity of the secretory granule membrane in the exocrine pancreas. The enzyme is also found in the granule content and in pancreatic secretions. Immunocytochemical localization confirmed its location in the granule membrane and in the acinar cell apical plasma membrane. In the endoplasmic reticulum and Golgi, membrane dipeptidase was strictly membrane-bound. There was no membrane dipeptidase in duct cells. The release of membrane dipeptidase from the membrane starts in the immature granule. To identify the mechanism responsible for its release, secretions were collected from cannulated conscious pig under basal conditions and atropine perfusion. The latter treatment caused complete inhibition of protein secretion but had a negligible effect on membrane dipeptidase activity in the secretions. In secretions, membrane dipeptidase partitioned into the detergent-rich phase on phase separation in Triton X-114, whereas treatment with bacterial phosphatidylinositol-specific phospholipase C caused the peptidase to partition into the aqueous phase, indicating that the secreted enzyme could come from shedding of membrane fragments at the apical surface or via the action of a previously characterized phospholipase A activity.

scholarly journals Adjustment of Hydrophobic Properties of Cellulose Materials

Polymers ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1241
Author(s):  
Michael Ioelovich
Keyword(s):  
Ester Group ◽  
Hydroxyl Groups ◽  
Chemical Methods ◽  
Hydrophobic Properties ◽  
Cellulose Esters ◽  
Cellulose Modification ◽  
Polar Groups ◽  
Polar Substituents ◽  
Hydrolysis Of ◽  
Cellulose Materials

In this study, physicochemical and chemical methods of cellulose modification were used to increase the hydrophobicity of this natural semicrystalline biopolymer. It has been shown that acid hydrolysis of the initial cellulose increases its crystallinity, which improves hydrophobicity, but only to a small extent. A more significant hydrophobization effect was observed after chemical modification by esterification, when polar hydroxyl groups of cellulose were replaced by non-polar substituents. The esterification process was accompanied by the disruption of the crystalline structure of cellulose and its transformation into the mesomorphous structure of cellulose esters. It was found that the replacement of cellulose hydroxyls with ester groups leads to a significant increase in the hydrophobicity of the resulting polymer. Moreover, the increase of the number of non-polar groups in the ester substituent contributes to rise in hydrophobicity of cellulose derivative. Depending on the type of ester group, the hydrophobicity increased in the following order: acetate < propionate < butyrate. Therefore, tributyrate cellulose (TBC) demonstrated the most hydrophobicity among all studied samples. In addition, the mixed ester, triacetobutyrate cellulose (TAB), also showed a sufficiently high hydrophobicity. The promising performance properties of hydrophobic cellulose esters, TBC and TAB, were also demonstrated.

Partial purification of a boar sperm membrane protein inducing sperm agglutinating antibody

1990 ◽  
Vol 10 (2) ◽  
pp. 131-139
Author(s):  
Oyewole Adeyemo ◽  
E. O. Okegbile ◽  
O. O. Olorunsogo
Keyword(s):  
Molecular Weight ◽  
Membrane Protein ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Partial Purification ◽  
Boar Sperm ◽  
Sperm Membrane ◽  
Triton X

For the development of immunological contraception, attention is being concentrated on the possibility of using a sperm membrane antigen. Boar sperm membrane was extracted with triton-X 100 and fractionated by Sephadex G-150 column chromatography. The glycosylated and nonglycosylated portions of protein peaks from the gel filtration were obtained by fractionating on concanavalin A-Sepharose and eluting the bound protein with 0.3 M methyl mannoside. A glycosylated fraction was found to induce sperm agglutinating antibodies in rabbit. The partially purified protein has a molecular weight of 30 kilodaltons, as determined by sodium dodecyl polyaccyrlamide gel electrophoresis. Further work is planned on the histochemical determination of the origin of this protein and species cross-activity of the antibody.

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