Assay of enzymes of lipid metabolism with colored and fluorescent derivatives of natural lipids.
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Animals↗
Biomedical subjects
B Shafit-Zagardo
Publications and source records attributed to B Shafit-Zagardo.
Assignment of the gene for acid beta-glucosidase to human chromosome 1.
The structural gene for human acid beta-glucosidase (GBA) has been assigned to chromosome 1 using somatic cell hybridization techniques for gene mapping. The human enzyme was detected in mouse RAG cell-human fibroblast cell hybrids by a sensitive double antibody immunoprecipitation assay using a mouse antihuman GBA antibody. No cross-reactivity between mouse beta-glucosidase and human GBA or neutral beta-glucosidase (GBN) was observed. Fifty-two primary, secondary, and tertiary manmouse hybrid lines, derived from three separate fusion experiments, were analyzed for human GBA and enzyme markers for the human chromosomes. Without exception, the presence of human GBA in these hybrid clones was correlated with the presence of human chromosome 1 or its enzymatic markers, phosphoglucomutase 1 (PGM1), and fumarate hydratase (FH). All other human chromosomes were eliminated by the independent segregation of GBA and their respective enzyme markers and/or chromosomes. Using a RAG X human fibroblast line with a mouse-human rearrangement of human chromosome 1, the locus for GBA was limited to the region 1p11 to 1qter.
Animals↗
Electrophoretic separation of neutral and acid beta-glucosidase isozymes in human tissues.
An electrophoretic system using cellulose acetate has been developed for the resolution of beta-glucosidase isozymes (beta-D-glucoside glucohydrolase, EC 3.2.1.21 and D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45) in human tissue homogenates. Electrophoresis of homogenates from normal and Type 1 Gaucher disease tissues revealed two fluorescent bands of beta-glucosidase activity which corresponded to the acid and neutral isozymes separated by concanavalin A-Sepharose chromatography. The acid isozyme has only beta-glucosidase activity, whereas the neutral isozyme also exhibited alpha-L-arabinosidase (alpha-L-arabinofuranoside arabinofuranohydrolase, EC 3.2.1.55), beta-D-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) and beta-D-xylosidase (1,4-beta-D-xylan xylohydrolase, EC 3.2.1.37) activities, using the appropriate 4-methylumbelliferyl glycoside. In homogenates of cultured skin fibroblasts, only the acid isozyme was observed which co-electrophoresed with the acidic activity in other tissue homogenates. The acidic activity in tissue and fibroblast homogenates from Type 1 Gaucher disease appeared to co-electrophorese with the acid isozyme in normal tissues, but had markedly reduced activity.
Brain↗
Transcription of the KpnI families of long interspersed DNAs in human cells.
The mammalian genome contains a variety of interspersed repetitive sequences of unknown function. It has, however, been suggested that interspersed repetitive sequences and their RNA transcripts are involved in the coordinate regulation of gene expression. Two major families of interspersed sequences in primates are the so-called Alu and KpnI families. Members of the KpnI families range in length from 1.2 to over 6 kilobases (kb). They exist in generally clustered arrangements, in 6 X 10(4) to 10(5) copies per diploid genome. Something is known of the arrangements of KpnI family sequences near human structural genes, but there has been no information on transcription of the sequences. We report here that the KpnI sequences are transcribed in HeLa cells by RNA polymerase II into abundant and heterogeneous species of RNA. The transcripts range in length from about 200 bases to over 5 kb, and are found predominantly in the non-polyadenylated fraction of the nuclear RNA. Transcripts homologous to both of the complementary strands of the KpnI sequences are present, but with a strong bias towards one strand.
Base Sequence↗