Document Detail


The effect of carbohydrate removal on stability and activity of saposin B.
MedLine Citation:
PMID:  8099782     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Saposin B is involved in the hydrolysis of sulfatides, GM1 ganglioside, globotriaosylceramide, and several other sphingolipids and glycerolipids by lysosomal hydrolases. Saposin B is one of four small glycoproteins (saposins) derived from prosaposin. The carbohydrate chain of saposin B was removed and deglycosylated saposin B was characterized and compared with native saposin B. Deglycosylated saposin B stimulated the enzymatic hydrolysis of ganglioside GM1 by acid beta-galactosidase and sulfatide by arylsulfatase A to the same extent as native saposin B. In addition deglycosylated saposin B bound sulfatide and GM1 ganglioside identical to native saposin B. The stability of native saposin B to proteolytic digestion was unchanged by deglycosylation. Neither native saposin B nor deglycosylated saposin B were hydrolyzed by trypsin, endoproteinase Glu-C (V-8), chymotrypsin, or a mixture of acid proteases isolated from human testis. Unlike its effect on metabolic stability, the carbohydrate chain appears to affect folding of saposin B. When native and deglycosylated saposin B were reduced under denaturing conditions and refolded under identical conditions examination of the refolded products indicated that each protein was refolded in a qualitatively different way. A human mutation in saposin B-deficient metachromatic leukodystrophy, in which its glycosylation site is eliminated, has been reported. Our observations suggest that instability of the mutated saposin B is not due to the absence of a protective effect of the carbohydrate chain on proteolysis, but is likely due to aberrant folding resulting from the absence of a carbohydrate chain.
Authors:
M Hiraiwa; S Soeda; B M Martin; A L Fluharty; Y Hirabayashi; J S O'Brien; Y Kishimoto
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Archives of biochemistry and biophysics     Volume:  303     ISSN:  0003-9861     ISO Abbreviation:  Arch. Biochem. Biophys.     Publication Date:  1993 Jun 
Date Detail:
Created Date:  1993-07-09     Completed Date:  1993-07-09     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0372430     Medline TA:  Arch Biochem Biophys     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  326-31     Citation Subset:  IM    
Affiliation:
Department of Neurosciences, University of California, San Diego, La Jolla 92093-0634.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Carbohydrates / chemistry*
Cerebroside-Sulfatase / metabolism
Chromatography, High Pressure Liquid
Drug Stability
Electrophoresis, Polyacrylamide Gel
G(M1) Ganglioside / metabolism
Gaucher Disease / metabolism
Glycoproteins / chemistry*,  metabolism,  pharmacology
Glycosylation
Humans
Hydrolysis
Saposins
Sphingolipid Activator Proteins
Spleen / chemistry
Structure-Activity Relationship
Sulfoglycosphingolipids / metabolism
beta-Galactosidase / metabolism
Grant Support
ID/Acronym/Agency:
NS-08682/NS/NINDS NIH HHS; NS-13559/NS/NINDS NIH HHS; NS-22655/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Carbohydrates; 0/Glycoproteins; 0/PSAP protein, human; 0/Saposins; 0/Sphingolipid Activator Proteins; 0/Sulfoglycosphingolipids; 37758-47-7/G(M1) Ganglioside; EC 3.1.6.8/Cerebroside-Sulfatase; EC 3.2.1.23/beta-Galactosidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Biosynthetic and structural studies on Thy-1 in a rat neuronal tumor cell line.
Next Document:  Identification and perturbation of mutant human fibroblasts based on measurements of methylmalonic a...