1,720,973 research outputs found
Identification of beta-N-acetylhexosaminidase A in mouse tissue with the fluorigenic substrate 4-methylumbelliferyl-beta-N-acetylglucosamine 6-sulphate.
No full textbeta-N-Acetylhexosaminidase from mouse tissue was separated into its constituent isoenzymes on DEAE-cellulose and its activity was monitored with 4-methylumbelliferyl-beta-N-acetylglucosamine and 4-methylumbelliferyl-beta-N-acetylglucosamine 6-sulphate. Forms corresponding to the human isoenzymes A (acidic), B (basic) and an 'intermediate' form were present in mouse liver and spleen, whereas in kidney the B and 'intermediate' forms predominated, with A present only as a minor component. In brain the 'intermediate', A and C activities were detected. Testis had predominantly A activity, whereas epididymis, the tissue with the highest specific activity of beta-N-acetylhexosaminidase, had an abundance of the 'intermediate' form, but was almost entirely lacking in the A form
Influence of cell differentiation and protein kinase C activation on sub-cellular distribution of beta-N-acetylhexosaminidases of HL 60 cells.
No full textThere have been several accounts regarding the alterations of the lysosomal enzyme beta-N-acetylhexosaminidase in human leukaemic cells. In addition to Hex A (alpha beta) and Hex B (beta beta) forms, leukaemic cells contain a third isoenzyme displaying many characteristics in common with Hex S, the alpha alpha dimer representing the residual activity in patients with Sandhoff's disease. In the human leukaemic cell line HL 60, A (alpha beta) and S (alpha alpha) are the most abundant forms. Sub-cellular fractionation of HL 60 cells showed that both A and S forms were present in the lysosomal and post-lysosomal fractions, however, a proportion of activity was found to be associated with the plasma membrane. The phorbol ester 12-O-tetra-decanoylphorbol-13-acetate (TPA) exerts complex effects on the physiology of HL 60 cells, leading to cell differentiation along the macrophage pathway and including activation of Protein Kinase C (PKC). In order to assess the extent to which cell differentiation and PKC activation plays a role in modulating the expression of hexosaminidase during cell differentiation, we treated HL 60 cells with TPA and in parallel with the more specific activator of PKC, 1-oleoyl-2-acetyl diglycerol (OAG) which does not cause cell differentiation. We observed that 24 h exposure of HL 60 cells to TPA or OAG produced significant modification of the hexosaminidase isoenzyme pattern of HL 60 cells. The most remarkable effect was seen in both cases in the plasma membrane fraction. Taken together, our results suggest a correlation between hexosaminidase expression and kinase(s) activation
Beta-N-acetylhexosaminidase in the splen of a patient with hairy-cell leukaemia.
No full texthe spleen from a patient with hairy-cell leukaemia had beta-N-acetylhexosaminidase activity that could be resolved into three isoenzymes by chromatography on phenyl boronate agarose. Two of these were the major forms, A and B, found in normal tissues but, in addition, there was an 'extra' form that accounted for 15% of total activity. The 'extra' form hydrolysed the synthetic substrate 4-methylumbelliferyl-beta-N-acetylglucosamine 6-sulphate, indicating the presence of alpha-subunits. It was more acidic than A, was less heat-stable and showed no generation of B on denaturation under a variety of conditions. These findings and the immunoblot (Western blotting) analysis demonstrate that the 'extra' form is entirely composed of alpha-subunits, and most closely resembles S, the residual activity in Sandhoff's disease
Cloning and sequence analysis of cDNA encoding the alpha-sub-unit of mouse beta-N-acetylhexosaminidase and comparison with the human enzyme
No full textcDNAs encoding the mouse beta-N-acetylhexosaminidase alpha-subunit were isolated from a mouse testis library. The longest of these (1.7 kb) was sequenced and showed 83% similarity with the human alpha-subunit cDNA sequence. The 5' end of the coding sequence was obtained from a genomic DNA clone. Alignment of the human and mouse sequences showed that all three putative N-glycosylation sites are conserved, but that the mouse alpha-subunit has an additional site towards the C-terminus. All eight cysteines in the human sequence are conserved in the mouse. There are an additional two cysteines in the mouse alpha-subunit signal peptide. All amino acids affected in Tay-Sachs-disease mutations are conserved in the mouse
Cloning and sequence analysis of a cDNA clone coding for the mouse GM2 activator protein.
No full textA cDNA (1.1 kb) containing the complete coding sequence for the mouse G(M2) activator protein was isolated from a mouse macrophage library using a cDNA for the human protein as a probe. There was a single ATG located 12 bp from the 5' end of the cDNA clone followed by an open reading frame of 579 bp. Northern blot analysis of mouse macrophage RNA showed that there was a single band with a mobility corresponding to a size of 2.3 kb. We deduce from this that the mouse mRNA, in common with the mRNA for the human G(M2) activator protein, has a long 3' untranslated sequence of approx. 1.7 kb. Alignment of the mouse and human deduced amino acid sequences showed 68% identity overall and 75% identity for the sequence on the C-terminal side of the first 31 residues, which in the human G(M2) activator protein contains the signal peptide. Hydropathicity plots showed great similarity between the mouse and human sequences even in regions of low sequence similarity. There is a single N-glycosylation site in the mouse G(M2) activator protein sequence (Asn151-Phe-Thr) which differs in its location from the single site reported in the human G(M2) activator protein sequence (Asn63-Val-Thr)
4-METHYLUMBELLIFERYL-ß-N-ACETYLGLUCOSAMINE-6-SULPHATE FOR CARRIER DETECTION OF TAY-SACHS DISEASE
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Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
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