scholarly journals Turnover of β-galactosidase in fibroblasts from patients with genetically different types of β-galactosidase deficiency

1981 ◽  
Vol 200 (1) ◽  
pp. 143-151 ◽  
Author(s):  
O P Van Diggelen ◽  
A W Schram ◽  
M L Sinnott ◽  
P J Smith ◽  
D Robinson ◽  
...  
Keyword(s):  
Hydrolytic Activity ◽  
Cellular Protein ◽  
Turnover Time ◽  
Galactosidase Activity ◽  
Normal Amount ◽  
Gm1 Gangliosidosis ◽  
Different Types ◽  
Suicide Substrate ◽  
Mutant Cells ◽  
Beta Galactosidase

The turnover of lysosomal beta-galactosidase was studied in fibroblast cultures from patients with Gm1-gangliosidosis and combined beta-galactosidase and neuraminidase deficiency, which had 5-10% residual beta-galactosidase activity. beta-Galactosidase was specifically inactivated with the suicide substrate beta-D-galactopyranosylmethyl-p-nitro-phenyltriazene (beta-Gal-MNT) and from the subsequent restoration of enzyme activity in cell cultures turnover times were calculated. By using [3H]beta-Gal-MNT, the hydrolytic activity per molecule of beta-galactosidase was determined. 3H-labelled beta-D-galactopyranosylmethylamine, the precursor of [3H]beta-gal-MNT, was obtained by Raney-nickel-catalysed exchange with 3H2O. The rate of synthesis of beta-galactosidase in normal and all mutant cells tested was found to be 0.4-0.5 pmol/day per mg of cellular protein. The GM1-gangliosidosis cells tested contain the normal amount of 0.5 pmol of beta-galactosidase/mg of protein with a normal turnover time of about 10 days, but only 10% of beta-galactosidase activity per enzyme molecule. Cells with combined beta-galactosidase and neuraminidase deficiency contain only 0.3 pmol of beta-galactosidase/mg of protein with a decreased turnover time of 1 day and normal hydrolytic properties (200 nmol of 4-methylumbelliferyl galactoside/h pmol of beta-galactosidase).

scholarly journals GM1 - type 1 glanglio sido sis: anatomo-clinic study of a case

1987 ◽  
Vol 45 (1) ◽  
pp. 60-66
Author(s):  
Ligia Maria Barbosa-Coutinho ◽  
Beatriz Maria Assis-Brasil ◽  
Maria de Lourdes Drachler ◽  
Newra Tellechea Rotta ◽  
Roberto Giuliani
Keyword(s):  
Respiratory Distress ◽  
Histological Examination ◽  
Fatal Outcome ◽  
Disease Diagnosis ◽  
Galactosidase Activity ◽  
Severe Respiratory Distress ◽  
Gm1 Gangliosidosis ◽  
Beta Galactosidase ◽  
Clinic Study

The observation of generalized GM1 gangliosidosis type 1 (Norman-Landing disease) is reported. The case is typical, featuring all the main clinical and biological signs of the disease. Diagnosis was established by the demonstration of a severe deficit in beta-galactosidase activity in leucocytes, by the demonstration of oligosacarides in the urine, and by the histological examination after the fatal outcome before the age of two with severe respiratory distress.

Leukocyte β-Galactosidase Activity in GM1-Gangliosidosis

PEDIATRICS ◽  
1972 ◽  
Vol 50 (3) ◽  
pp. 502-503
Author(s):  
Elisabeth Young ◽  
R. B. Ellis ◽  
A. D. Patrick
Keyword(s):  
Citric Acid ◽  
Recent Article ◽  
Galactosidase Activity ◽  
Alternative Substrate ◽  
Gm1 Gangliosidosis ◽  
Acid Buffer ◽  
Assay Medium ◽  
Routine Service ◽  
Citric Acid Buffer ◽  
Beta Galactosidase

The recent article by Singer et al. entitled "Leukocyte beta-Galactosidase Activity in the Diagnosis of Generalized GM1-Gangliosidosis," prompts us to report the results of two years of routine service assay of this enzyme in leukocytes using the alternative substrate, 4-methyl-umbelliferyl β-D-galactopyranoside. The β-galactosidase assay medium contained McIlvaine phosphate-citric acid buffer, pH, 4.1 (40µl); 4-methylumbelliferyl β-D-galactopyranoside (1 mM in buffer, 150µl) and leukocyte extract in 0.2 MKCl (10µl). After incubation for 15 minutes at 30°C, the reaction was stopped by the addition of 0.25 M glycine-NaOH, pH, 10.4 (1 ml) and the fluorescence measured.

scholarly journals Programmed cell senescence in the mouse developing spinal cord and notochord

2020 ◽  
Author(s):  
Jorge Antolio Domínguez-Bautista ◽  
Pilar Sarah Acevo-Rodríguez ◽  
Susana Castro-Obregón
Keyword(s):  
Spinal Cord ◽  
Nervous System ◽  
Cellular Senescence ◽  
Cell Senescence ◽  
Surrounding Tissue ◽  
Galactosidase Activity ◽  
Mouse Development ◽  
Different Types ◽  
Beta Galactosidase ◽  
Developing Spinal Cord

AbstractProgrammed cell senescence is a cellular process that seems to contribute to morphogenesis during embryo development, in addition to cell proliferation, migration, differentiation and programmed cell death, and has been observed in evolutionary distant organisms like mammals, amphibians and fish. Programmed cell senescence is a phenotype similar to stress-induced cellular senescence, characterized by the expression of cell cycle inhibitors such as CDKN1A/p21, increased activity of a lysosomal enzyme with beta-galactosidase activity (coined senescence-associated beta-galactosidase) and, most importantly, secretion of growth factors, interleukins, chemokines, metalloproteases, etc., collectively known as a senescent-associated secretory phenotype that instructs surrounding tissue. How wide is the distribution of programmed cell senescence during mouse development and its specific mechanisms to shape the embryo are still poorly understood. Here, we investigated whether markers of programmed cell senescence are found in the developing mouse spinal cord and notochord. We found discrete areas and developmental windows with high senescence-associated beta galactosidase in both spinal cord and notochord; expression of CDKN1A/p21 was documented in epithelial cells of the spinal cord and the notochord. Treatment of mice embryos developed ex-utero in the presence of the senolytic ABT-263 resulted in decrease senescence-associated beta-galactosidase activity and number of motoneurons. Our data suggest that several cell types undergo programmed cell senescence in developing spinal cord and notochord contributing to morphogenesis.Contribution to the Field StatementCellular senescence is a state in which cells no longer divide but have the remarkable ability to secrete signaling molecules that alter the tissue where they reside. In adults, this state is typically induced by stress situations that cause DNA damage so cells with altered genome do not multiply. Senescent cells also form when a tissue is injured; they help to regenerate damaged tissue and contribute to wound healing. Phagocytic cells eliminate them when their function is done, having a transient existence. During vertebrate development some cells acquire a very similar phenotype, coined programmed cell senescence, and interestingly they have been found in regions that organize the pattern of development of some organs. How wide is the distribution of programmed cell senescence during development and how they help to shape the embryo are still poorly understood. We discovered in mice embryos different types of cells with senescent features located in particular regions of the developing nervous system: where motoneurons form and in a region that secrete molecules that instruct the embryo where different types of neurons will be created. We propose that programed cell senescence contributes to the morphogenesis of the nervous system.

LEUKOCYTE BETA-GALACTOSIDASE ACTIVITY IN THE DIAGNOSIS OF GENERALIZED GM, GANGLIOSIDOSIS

PEDIATRICS ◽  
1972 ◽  
Vol 49 (3) ◽  
pp. 352-361
Author(s):  
Harvey S. Singer ◽  
George A. Nankervis ◽  
Irwin A. Schafer
Keyword(s):  
Enzyme Activity ◽  
Autosomal Recessive ◽  
Gm1 Ganglioside ◽  
Galactosidase Activity ◽  
Inherited Disease ◽  
Gm1 Gangliosidosis ◽  
Storage Diseases ◽  
Beta Galactosidase ◽  
Normal Controls ◽  
Test Tissue

GM1 generalized gangliosidosis is an autosomal recessive inherited disease, characterized by the storage of GM1 ganglioside in brain and visceral tissues secondary to a deficiency of the enzyme β-galactosidase. The enzyme deficiency found in brain, liver, and cultured flbroblasts is also present in the leukocyte. Using leukocytes as the test tissue two GM1 gangliosidosis putients were clearly identified from patients with clinically similar storage diseases. The enzyme activity in obligate heterozygotes showed intermediate levels between their affected children and normal controls, suggesting that heterozygotes may be identified using the leukocyte as the test tissue.

Overlapping reading frames at the LYS5 locus in the yeast Yarrowia lipolytica

1990 ◽  
Vol 10 (9) ◽  
pp. 4795-4806
Author(s):  
J W Xuan ◽  
P Fournier ◽  
N Declerck ◽  
M Chasles ◽  
C Gaillardin
Keyword(s):  
Yarrowia Lipolytica ◽  
Stop Codon ◽  
Open Reading Frames ◽  
Yeast Cells ◽  
Lacz Gene ◽  
Galactosidase Activity ◽  
Phenotypic Effect ◽  
Frame Fusion ◽  
Beta Galactosidase ◽  
Reading Frames

Mutants affected at the LYS5 locus of Yarrowia lipolytica lack detectable dehydrogenase (SDH) activity. The LYS5 gene has previously been cloned, and we present here the sequence of the 2.5-kilobase-pair (kb) DNA fragment complementing the lys5 mutation. Two large antiparallel open reading frames (ORF1 and ORF2) were observed, flanked by potential transcription signals. Both ORFs appear to be transcribed, but several lines of evidence suggest that only ORF2 is translated and encodes SDH. (i) The global amino acid compositions of Saccharomyces cerevisiae SDH and of the putative ORF2 product are similar and that of ORF1 is dissimilar. (ii) An in-frame translational fusion of ORF2 with the Escherichia coli lacZ gene was introduced into yeast cells and resulted in a beta-galactosidase activity regulated similarly to SDH; no beta-galactosidase activity was obtained with an in-frame fusion of ORF1 with lacZ. (iii) The introduction of a stop codon at the beginning of ORF2 prevented SDH expression in yeast cells, whereas no phenotypic effect was observed when ORF1 translation was blocked.

Beta-galactosidase deficiencies and novel GLB1 mutations in three Chinese patients with Morquio B disease or GM1 gangliosidosis

2012 ◽  
Vol 8 (4) ◽  
pp. 359-362 ◽  
Author(s):  
Hong-Lin Lei ◽  
Jun Ye ◽  
Wen-Juan Qiu ◽  
Hui-Wen Zhang ◽  
Lian-Shu Han ◽  
...  
Keyword(s):  
Chinese Patients ◽  
Gm1 Gangliosidosis ◽  
Beta Galactosidase
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