scholarly journals Renal dipeptidase is one of the membrane proteins released by phosphatidylinositol-specific phospholipase C.

1987 ◽  
Vol 244 (2) ◽  
pp. 465-469 ◽  
Author(s):  
N M Hooper ◽  
M G Low ◽  
A J Turner
Keyword(s):  
Staphylococcus Aureus ◽  
Alkaline Phosphatase ◽  
Phase Separation ◽  
Bacillus Thuringiensis ◽  
Bacillus Cereus ◽  
Phospholipase C ◽  
Membrane Fraction ◽  
Similar Extent ◽  
Pig Kidney ◽  
Triton X

Renal dipeptidase (dehydropeptidase-I, EC 3.4.13.11) was released from pig kidney membrane preparations by treatment with phosphatidylinositol-specific phospholipase C from Staphylococcus aureus and Bacillus thuringiensis and a phospholipase C preparation from Bacillus cereus to a similar extent as alkaline phosphatase. Endopeptidase-24.11 and aminopeptidase N were not released by this treatment. After treatment of the membrane fraction with the S. aureus phospholipase C the dipeptidase was converted from an amphipathic to a hydrophilic form, as deduced from phase-separation experiments in Triton X-114. It is concluded that renal dipeptidase is anchored to the microvillar membrane by covalently attached phosphatidylinositol.

Incorporation of human liver and placental alkaline phosphatases into liposomes and membranes is via phosphatidylinositol

1990 ◽  
Vol 68 (9) ◽  
pp. 1112-1118 ◽  
Author(s):  
Lee Kihn ◽  
Dorothy Rutkowski ◽  
Robert A. Stinson
Keyword(s):  
Alkaline Phosphatase ◽  
Phospholipase C ◽  
Human Liver ◽  
Red Cell ◽  
Osteosarcoma Cells ◽  
Alkaline Phosphatases ◽  
Membrane Anchor ◽  
Gradient Gel Electrophoresis ◽  
Triton X ◽  
Phosphatidylinositol Phospholipase C

As assessed by incorporation into liposomes and by adsorption to octyl-Sepharose, the integrity of the membrane anchor for the purified tetrameric forms of alkaline phosphatase from human liver and placenta was intact. Any treatment that resulted in a dimeric enzyme precluded incorporation and adsorption. An intact anchor also allowed incorporation into red cell ghosts. The addition of hydrophobic proteins inhibited incorporation into liposomes to varying degrees. Alkaline phosphatase was 100% releasable from liposomes and red cell ghosts by a phospholipase C specific for phosphatidylinositol. There was no appreciable difference in the rates of release of placental and liver alkaline phosphatases, although both were approximately 250 × slower in liposomes and 100 × slower in red cell ghosts than the enzyme's release from a suspension of cultured osteosarcoma cells. Both enzymes were released by phosphatidylinositol phospholipase C as dimers and would not reincorporate or adsorb to octyl-Sepharose. However, the enzyme incorporated, resolubilized by Triton X-100, and cleansed of the detergent by butanol treatment was tetrameric by gradient gel electrophoresis, was hydrophobic, and could reincorporate into fresh liposomes. A monoclonal antibody to liver alkaline phosphatase inhibited the enzyme's incorporation into liposomes, and abolished its release from liposomes and its conversion to dimers by phosphatidylinositol phospholipase C.Key words: alkaline phosphatase, liposome, phosphatidylinositol, membrane anchor.

scholarly journals Sequence Analysis of Inducible Prophage phIS3501 Integrated into the Haemolysin II Gene of Bacillus thuringiensis var israelensis ATCC35646

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Bouziane Moumen ◽  
Christophe Nguen-The ◽  
Alexei Sorokin
Keyword(s):  
Staphylococcus Aureus ◽  
Sequence Analysis ◽  
Bacillus Thuringiensis ◽  
Bacillus Cereus ◽  
Genomic Organization ◽  
Food Poisoning ◽  
Complete Sequence ◽  
Bacterial Host ◽  
Regulation Of Synthesis ◽  
Selection For

Diarrheic food poisoning by bacteria of the Bacillus cereus group is mostly due to several toxins encoded in the genomes. One of them, cytotoxin K, was recently identified as responsible for severe necrotic syndromes. Cytotoxin K is similar to a class of proteins encoded by genes usually annotated as haemolysin II (hlyII) in the majority of genomes of the B. cereus group. The partially sequenced genome of Bacillus thuringiensis var israelensis ATCC35646 contains several potentially induced prophages, one of them integrated into the hlyII gene. We determined the complete sequence and established the genomic organization of this prophage-designated phIS3501. During induction of excision of this prophage with mitomycin C, intact hlyII gene is formed, thus providing to cells a genetic ability to synthesize the active toxin. Therefore, this prophage, upon its excision, can be implicated in the regulation of synthesis of the active toxin and thus in the virulence of bacterial host. A generality of selection for such systems in bacterial pathogens is indicated by the similarity of this genetic arrangement to that of Staphylococcus aureus  β-haemolysin.

scholarly journals Release of alkaline phosphatase from membranes by a phosphatidylinositol-specific phospholipase C.

1977 ◽  
Vol 167 (1) ◽  
pp. 281-284 ◽  
Author(s):  
M G Low ◽  
J B Finean
Keyword(s):  
Staphylococcus Aureus ◽  
Alkaline Phosphatase ◽  
Phospholipase C ◽  
Mammalian Species ◽  
Salt Solutions ◽  
Total Alkaline Phosphatase ◽  
Total Alkaline Phosphatase Activity ◽  
Wide Range ◽  
Releasing Effect ◽  
Total Alkaline

Purified phosphatidylinositol-specific phospholipase C from Staphylococcus aureus released a substantial proportion of the total alkaline phosphatase activity from a wide range of tissues from several mammalian species. Co-purification of the phospholipase C and alkaline phosphatase-releasing activities and the inhibition of both these activities by iso-osmotic salt solutions suggested that the releasing effect was unlikely to be due to a contaminant.

Effect of phospholipase C on high-molecular-mass alkaline phosphatase in serum.

1986 ◽  
Vol 32 (8) ◽  
pp. 1503-1505 ◽  
Author(s):  
E Sykes ◽  
F L Kiechle ◽  
E Epstein
Keyword(s):  
Alkaline Phosphatase ◽  
Bacillus Cereus ◽  
Molecular Mass ◽  
Phospholipase C ◽  
Clostridium Perfringens ◽  
Alkaline Phosphatase Activity ◽  
Plasma Membranes ◽  
High Molecular Mass ◽  
Serum Samples ◽  
Liver Membrane

Abstract Electrophoresis of some serum samples on polyacrylamide gel, followed by staining for alkaline phosphatase (EC 3.1.3.1), produces a band of activity at the gel origin. This high-Mr band consists of liver membrane fragments containing alkaline phosphatase and other enzymes. Alkaline phosphatase is closely associated with phosphatidylinositol in liver plasma membranes, and we have found that phospholipase C (EC 3.1.4.3) from Bacillus cereus, known to possess some phosphatidylinositol specificity, was able to release liver alkaline phosphatase from the high-Mr band. Two preparations of phospholipase C from Clostridium perfringens, however, which has no phosphatidylinositol specificity, had no effect on the alkaline phosphatase activity in the high-Mr band.

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.

Endogenous glycosylphosphatidylinositol-specific phospholipase C releases renal dipeptidase from kidney proximal tubules in vitro

2001 ◽  
Vol 353 (2) ◽  
pp. 339-344 ◽  
Author(s):  
Sung Wook PARK ◽  
Kyong CHOI ◽  
Cheol KIM ◽  
Hwang Hee Blaise LEE ◽  
Nigel M. HOOPER ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Phase Separation ◽  
Phospholipase C ◽  
Aqueous Phase ◽  
Proximal Tubules ◽  
Gpi Anchor ◽  
Ph Optimum ◽  
Hydrophilic Nature ◽  
Triton X

Spontaneous enzymic release of renal dipeptidase (RDPase; EC 3.4.13.19), a glycosylphosphatidylinositol (GPI)-linked ectoenzyme, was observed in vitro during incubation of porcine proximal tubules at 37°C. Triton X-114 phase separation of the released RDPase showed that the majority of the enzyme activity partitioned into the aqueous phase, indicating its hydrophilic nature. Immunoblot analyses using an antibody against the cross-reacting determinant (CRD) inositol 1,2-cyclic monophosphate, the epitope formed by phospholipase C (PLC) cleavage of the GPI anchor on a protein, detected the released RDPase. Reprobing the immunoblot with an anti-RDPase serum showed the RDPase band co-migrating with the CRD band. The release of RDPase from the proximal tubules was a Ca2+-dependent process and had a pH optimum of 9.0. These results indicate that RDPase is released from the proximal tubules by the action of a distinct endogenous GPI-specific PLC.

scholarly journals Biosynthesis of glycosylphosphatidylinositol-anchored human placental alkaline phosphatase: evidence for a phospholipase C-sensitive precursor and its post-attachment conversion into a phospholipase C-resistant form

1994 ◽  
Vol 301 (1) ◽  
pp. 205-209 ◽  
Author(s):  
Y W Wong ◽  
M G Low
Keyword(s):  
Alkaline Phosphatase ◽  
Phospholipase C ◽  
Hela Cells ◽  
Placental Alkaline Phosphatase ◽  
Gpi Anchor ◽  
Chase Period ◽  
Resistant Species ◽  
Resistant Form ◽  
Human Placental Alkaline Phosphatase ◽  
Triton X

Previous studies have shown that some cells (e.g. SKG3a) express human placental alkaline phosphatase (AP) in a form which can be released from the membrane by bacterial PtdIns-specific phospholipase C (PI-PLC) while others (e.g. HeLa) are relatively resistant to this enzyme. Chemical and enzymic degradation studies have suggested that the PI-PLC resistance of AP is due to inositol acylation of its glycosylphosphatidylinositol (GPI) anchor. In order to identify the biosynthetic origin of PI-PLC resistance we determined the PI-PLC sensitivity of AP in 35S-labelled cells (10 min pulse; 0-60 min chase) by Triton X-114 phase separation. At the beginning of the chase period, the majority of the AP synthesized was hydrophilic, indicating that it had not acquired a GPI anchor. The concentration of hydrophilic AP species decreased with a t1/2 of 30-60 min but was not processed to an endoglycosidase H-resistant species or secreted into the medium. In both SKG3a and HeLa cells all of the hydrophobic, GPI-anchored AP detectable at the beginning of the chase was PI-PLC sensitive. PI-PLC-resistant species of AP were only observed in HeLa cells and these only appeared after about 30 min. The delayed appearance of PI-PLC resistance was unexpected as previous studies have suggested that candidate GPI-anchor precursors are PI-PLC-resistant as a result of inositol acylation. This work reveals unanticipated complexities in the biosynthesis of AP and its GPI anchor.

Physiological actions of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis on KB III cells: Alkaline phosphatase release and growth inhibition

Toxicon ◽  
1985 ◽  
Vol 23 (1) ◽  
pp. 145-155 ◽  
Author(s):  
Hiroh Ikezawa ◽  
Takashi Kato ◽  
Shin-Ichi Ohta ◽  
Toshikatsu Nakabayashi ◽  
Yukari Iwata ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Bacillus Thuringiensis ◽  
Growth Inhibition ◽  
Phospholipase C
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