scholarly journals Isolation and characterization of an activator protein for the hydrolysis of ganglioside GM2 from the roe of striped mullet (Mugil cephalus)

1989 ◽  
Vol 260 (3) ◽  
pp. 777-783 ◽  
Author(s):  
R DeGasperi ◽  
Y T Li ◽  
S C Li
Keyword(s):  
Mugil Cephalus ◽  
Striped Mullet ◽  
Activator Protein ◽  
Isolation And Characterization ◽  
Mammalian Tissues ◽  
Hexosaminidase A ◽  
Ammonium Sulphate Fractionation ◽  
Gm2 Activator ◽  
Hydrolysis Of

After the revelation of the presence of ganglioside GM2 as the major ganglioside in the roe of striped mullet, Mugil cephalus [Li, Hirabayashi, DeGasperi, Yu, Ariga, Koerner & Li (1984) J. Biol. Chem. 259, 8980-8985], we have continued to investigate the catabolism of GM2 in this tissue. We have found that mullet roe contains a specific activator protein which stimulates the hydrolysis of GM2 carried out by the beta-hexosaminidase isolated from the same tissue. This activator has been purified by using conventional procedures including ammonium sulphate fractionation and chromatography on Sepharose 6B, DEAE-Sephadex A-50, octyl-Sepharose and Matrex Gel Blue A columns. This activator protein is also able to stimulate the hydrolysis of GM2 carried out by human beta-hexosaminidase A. Unlike human GM2-activator, the roe activator protein does not stimulate the hydrolysis of GgOse3Cer or GbOse4Cer. The molecular mass (18 kDa) of the roe activator protein was found to be similar to that of human GM2-activator; however, the pI (pH 4.1) was found to be lower than that of human GM2-activator. This is the first report on the presence of a GM2-activator protein in a source other than mammalian tissues.

Isolation and Characterization of an Anaerobic Bacterium, Eubacterium tarantellus sp.nov., Associated with Striped Mullet (Mugil cephalus) Mortality in Biscayne Bay, Florida

1977 ◽  
Vol 34 (3) ◽  
pp. 402-409 ◽  
Author(s):  
Lanny R. Udey ◽  
Elaine Young ◽  
Bennett Sallman
Keyword(s):  
Ictalurus Punctatus ◽  
Anaerobic Bacterium ◽  
Biochemical Characterization ◽  
Anaerobic Bacteria ◽  
Mugil Cephalus ◽  
Estuarine Fish ◽  
Biscayne Bay ◽  
Striped Mullet ◽  
Isolation And Characterization

An asporogenous, gram-positive, anaerobic bacterium was isolated in pure culture from the brains of numerous dead and moribund striped mullet (Mugil cephalus) from Biscayne Bay, Florida. Biochemical characterization confirmed that it was a new species and has been named Eubacterium tarantellus (ATCC 29255). All isolates produced lecithinase and deoxyribonuclease, were beta hemolytic, but only weakly fermented selected carbohydrates. This anaerobe was pathogenic for channel catfish (Ictalurus punctatus) but not for white mice or guinea pigs.More recently E. tarantellus has been isolated from 10 additional fish species. This and other anaerobic bacteria may be significant pathogens of estuarine fish. Key words: Anaerobic bacteria, Eubacterium, Mugil cephalus, and fish diseases.

Isolation and characterization of microsatellite loci in the striped mullet, Mugil cephalus

2005 ◽  
Vol 5 (2) ◽  
pp. 323-326 ◽  
Author(s):  
E. MIGGIANO ◽  
R. E. LYONS ◽  
Y. LI ◽  
L. M. DIERENS ◽  
D. CROSETTI ◽  
...  
Keyword(s):  
Microsatellite Loci ◽  
Mugil Cephalus ◽  
Striped Mullet ◽  
Isolation And Characterization

Isolation and Characterization of Calpain Activator Protein from Bovine Brain

2003 ◽  
pp. 99-105
Author(s):  
Edon Melloni ◽  
Mauro Michetti ◽  
Franca Salamino ◽  
Roberto Minafra ◽  
Bianca Sparatore ◽  
...  
Keyword(s):  
Bovine Brain ◽  
Activator Protein ◽  
Isolation And Characterization

Isolation and characterization of anti-inflammatory peptides derived from trypsin hydrolysis of microalgae protein (Synechococcus sp. VDW)

2019 ◽  
Vol 33 (4) ◽  
pp. 303-324 ◽  
Author(s):  
Rutairat Suttisuwan ◽  
Saranya Phunpruch ◽  
Tanatorn Saisavoey ◽  
Papassara Sangtanoo ◽  
Nuttha Thongchul ◽  
...  
Keyword(s):  
Isolation And Characterization ◽  
Trypsin Hydrolysis ◽  
Synechococcus Sp ◽  
Anti Inflammatory ◽  
Hydrolysis Of

scholarly journals Isolation and characterization of a γ-type phosphoinositide-specific phospholipase C (PLC-γ2)

1990 ◽  
Vol 269 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Y Homma ◽  
Y Emori ◽  
F Shibasaki ◽  
K Suzuki ◽  
T Takenawa
Keyword(s):  
Phospholipase C ◽  
Column Chromatography ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Isolation And Characterization ◽  
Delta Type ◽  
Hydrolysis Of ◽  
Cellulose Column

A novel bovine spleen phosphoinositide-specific phospholipase C (PLC) has been identified with respect to immunoreactivity with four independent antibodies against each of the PLC isoenzymes, and purified to near homogeneity by sequential column chromatography. Spleen contains three of the isoenzymes: two different gamma-types [gamma 1 and gamma 2, originally named as PLC-gamma [Rhee, Suh, Ryu & Lee (1989) Science 244, 546-550] and PLC-IV [Emori, Homma, Sorimachi, Kawasaki, Nakanishi, Suzuki & Takenawa (1989) J. Biol. Chem. 264, 21885-21890] respectively] and delta-type of the enzyme, but PLC-gamma 1 is separated from the PLC-gamma 2 pool by the first DEAE-cellulose column chromatography. Subsequently, PLC-delta is dissociated on the third heparin-Sepharose column chromatography. The purified enzyme has a molecular mass of 145 kDa on SDS/polyacrylamide-gel electrophoresis and a specific activity of 12.8 mumol/min per mg with phosphatidylinositol 4,5-bisphosphate as substrate. This enzyme activity is dependent on Ca2+ for hydrolysis of all these phosphoinositides. None of the other phospholipids examined could be its substrate at any concentration of Ca2+. The optimal pH of the enzyme is slightly acidic (pH 5.0-6.5).

scholarly journals Interaction of activating protein and surfactants with human liver hexosaminidase A and GM2 ganglioside

1980 ◽  
Vol 185 (3) ◽  
pp. 583-591 ◽  
Author(s):  
Peter Hechtman ◽  
Zarin Kachra
Keyword(s):  
Human Liver ◽  
Anionic Surfactants ◽  
Sodium Taurocholate ◽  
Ionic Surfactants ◽  
Enzymic Hydrolysis ◽  
Gm2 Ganglioside ◽  
Activator Protein ◽  
Hexosaminidase A ◽  
Triton X ◽  
Hydrolysis Of

The effects of surfactants on the human liver hexosaminidase A-catalysed hydrolysis of Gm2 ganglioside were assessed. Some non-ionic surfactants, including Triton X-100 and Cutscum, and some anionic surfactants, including sodium taurocholate, sodium dodecyl sulphate, phosphatidylinositol and N-dodecylsarcosinate, were able to replace the hexosaminidase A-activator protein [Hechtman (1977) Can. J. Biochem.55, 315–324; Hechtman & Leblanc (1977) Biochem. J.167, 693–701) and also stimulated the enzymic hydrolysis of substrate in the presence of saturating concentrations of activator. Other non-ionic surfactants, such as Tween 80, Brij 35 and Nonidet P40, and anionic surfactants, such as phosphatidylethanolamine, did not enhance enzymic hydrolysis of Gm2 ganglioside and inhibited hydrolysis in the presence of activator. The concentration of surfactants at which micelles form was determined by measurements of the minimum surface-tension values of reaction mixtures containing a series of concentrations of surfactant. In the case of Triton X-100, Cutscum, sodium taurocholate, N-dodecylsarcosinate and other surfactants the concentration range at which stimulation of enzymic activity occurs correlates well with the critical micellar concentration. None of the surfactants tested affected the rate of hexosaminidase A-catalysed hydrolysis of 4-methylumbelliferyl N-acetyl-β-d-glucopyranoside. Both activator and surfactants that stimulate hydrolysis of Gm2 ganglioside decrease the Km for Gm2 ganglioside. Inhibitory surfactants are competitive with the activator protein. Evidence for a direct interaction between surfactants and Gm2 ganglioside was obtained by comparing gel-filtration profiles of 3H-labelled GM2 ganglioside in the presence and absence of surfactants. The results are discussed in terms of a model wherein a mixed micelle of surfactant or activator and GM2 ganglioside is the preferred substrate for enzymic hydrolysis.

Substrate binding properties of the human liver hexosaminidase A activator protein

1985 ◽  
Vol 63 (8) ◽  
pp. 830-838 ◽  
Author(s):  
Peter Hechtman ◽  
Claudine Isaacs ◽  
Louise Smith-Jones
Keyword(s):  
Protein Complex ◽  
Human Liver ◽  
Substrate Binding ◽  
Enzyme Activation ◽  
Stable Form ◽  
Ph Optimum ◽  
Substrate Complex ◽  
Activator Protein ◽  
Hexosaminidase A ◽  
Hydrolysis Of

The human liver hexosaminidase A activator protein has been shown to bind to the substrate GM2 ganglioside by cosedimentation in sucrose density gradients. Among other proteins tested only serum albumin forms a GM2 ganglioside – protein complex. Both activator protein and albumin bind to the monomeric form of GM2 ganglioside and not to the micellar form of the substrate. The GM2 ganglioside – activator protein complex can be recovered in a stable form. Storage at various temperatures or incubation with monosaccharides or with detergent does not result in dissociation of the complex. GM2 ganglioside in the activator–substrate complex is exchangeable with exogenous GM2 ganglioside. Hexosaminidase A, prepared from human liver, hydrolyzes GM2 ganglioside in the activator–substrate complex as efficiently as GM2 ganglioside supplied exogenously. The activator – Gm2 ganglioside complex forms at pH 3.0 and exhibits an optimum similar to the pH optimum of hexosaminidase A catalyzed hydrolysis of GM2 ganglioside in the presence of the activator; however, the ability of the activator to stimulate enzymic hydrolysis of substrate is rapidly lost after heating at 75 °C, whereas its ability to bind substrate is increased. The sphingolipids cerebroside sulfate and sphingomyelin show little or no binding to the hexosaminidase A activator protein nor do they inhibit activation of hexosaminidase A catalyzed hydrolysis of GM2 ganglioside. By contrast GM1 ganglioside inhibits both substrate binding and enzyme activation.

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