| Synthesis and use of novel fluorescent glycosphingolipids for estimating beta-glucosidase activity in vitro in the absence of detergents and subtyping Gaucher disease variants following administration into intact cells. | |
| | |
MedLine Citation:
|
PMID: 8399329 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Two novel fluorescent glycolipids, LRO-glucosylceramide (LRO-GC) and LRO-trihexosylceramide (LRO-THC) were synthesized and utilized for estimating activities of the lysosomal, acid beta-glucosidase in cell extracts and intact skin fibroblasts, derived from normal individuals and patients with Gaucher disease subtypes. The uniqueness of the glycolipids is the fact that a fluorescent probe (lissamine rhodamine) is linked in a sulfonylamide linkage to the sphingosyl residue of the sphingolipid. Thus, the product of enzymatic hydrolysis, lissamine rhodamine sulfonylamido sphingosine (LRO-ceramide) cannot be further hydrolyzed and remains a metabolic end product. A unique property of LRO-GC as a substrate for the lysosomal, acid beta-glucosidase in vitro was the observation that enzymatic hydrolysis occurs in the absence of detergents and that hydrolytic rates are, in fact, reduced in the presence of Triton X-100 and/or sodium taurocholate. Also, both glycolipids penetrated the membrane of intact fibroblasts in the absence of serum and were hydrolyzed in lysosomes of the intact cells. The rates of intracellular hydrolysis decreased with the severity of the Gaucher disease subtypes. Using LRO-THC as substrate, the intracellular ratio of LRO-ceramide to LRO-glucosylceramide was an indicator for the specific GD-subtype. |
| | |
Authors:
|
V Agmon; S Cherbu; A Dagan; M Grace; G A Grabowski; S Gatt |
Related Documents
:
|
1666339 - Compartmentalization and actin binding properties of abp-50: the elongation factor-1 al... 8554919 - Localization of the ca(2+)-binding alpha-parvalbumin and its mrna in epiphyseal plate c... 8717139 - Benzylaminopurine induces phenocopies of floral meristem and organ identity mutants in ... 14724229 - Semaphorin3d guides retinal axons along the dorsoventral axis of the tectum. 17715149 - Cyanobacteriochrome tepixj of thermosynechococcus elongatus harbors phycoviolobilin as ... 1917849 - Glycine 100 in the dinitrogenase reductase of rhodospirillum rubrum is required for nit... 17102449 - Nursing intensity and patient classification in a patient information system. 3297219 - Computer-assisted instruction for medicine and nursing: sources and programs. 8516259 - Effects of public health nursing participation in selected academic courses on self-rep... |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
|
Title: Biochimica et biophysica acta Volume: 1170 ISSN: 0006-3002 ISO Abbreviation: Biochim. Biophys. Acta Publication Date: 1993 Sep |
Date Detail:
|
Created Date: 1993-11-01 Completed Date: 1993-11-01 Revised Date: 2007-11-14 |
Medline Journal Info:
|
Nlm Unique ID: 0217513 Medline TA: Biochim Biophys Acta Country: NETHERLANDS |
Other Details:
|
Languages: eng Pagination: 72-9 Citation Subset: IM |
Affiliation:
|
Department of Membrane Biochemistry and Neurochemistry, Hebrew University-Hadassah School of Medicine, Jerusalem, Israel. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Cells, Cultured Detergents Fluorescent Dyes / chemical synthesis* Gaucher Disease / classification, enzymology*, genetics Genotype Glucosylceramides / chemical synthesis Glycosphingolipids / chemical synthesis* Humans Point Mutation Rhodamines / chemical synthesis Skin / enzymology Trihexosylceramides / chemical synthesis beta-Glucosidase / analysis*, genetics |
| Grant Support | |
ID/Acronym/Agency:
|
DK 36729/DK/NIDDK NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/Detergents; 0/Fluorescent Dyes; 0/Glucosylceramides; 0/Glycosphingolipids; 0/Rhodamines; 0/Trihexosylceramides; 0/lissamine rhodamine sulfonylamidosphingosyl-trihexosylceramide; 0/lissamine rhodamine-sulfonylamidosphingosyl beta-glucoside; EC 3.2.1.21/beta-Glucosidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Early catalytic steps of Euglena gracilis chloroplast type II fatty acid synthase.
Next Document: Synthetic model peptides for apolipoproteins. II. Characterization of the discoidal complexes genera...