Document Detail


Biocytin and biotin uptake into NB2a neuroblastoma and C6 astrocytoma cells.
MedLine Citation:
PMID:  11792359     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Uptake of biocytin and biotin was investigated in cultured transformed variants of neuronal (NB2a neuroblastoma) and glial (C6 astrocytoma) CNS cells. NB2a cells took up both compounds but biocytin was transported more efficiently than biotin in the nanomolar concentration range. In NB2a cells a single transport mechanism was found for biocytin with different kinetic parameters in the presence of high extracellular Na+ (Km 0.4 microM, Vmax 20 pmol/min/mg), K+ (Km 1.7 microM, Vmax 32 pmol/min/mg), or choline+ (Km 0.1 microM, Vmax 5 pmol/min/mg). Two transport systems (Km1 17 microM, Vmax1 53 pmol/min/mg; Km2 314 microM, Vmax2 360 pmol/min/mg) were identified for biotin with only system 1 being Na+-dependent. Biocytin uptake was competitively inhibited by excess biotin but not vice versa. Inhibition studies with structural analogs indicated different specificities for biotin and biocytin uptake. Biocytin uptake into C6 cells was hardly detectable whereas biotin was taken up by diffusion (kD 0.6 microl/min/mg) and a single saturable mechanism (Km 70 microM, Vmax 119 pmol/min/mg) at high extracellular Na+. High extracellular K+ enhanced biotin diffusion into C6 cells. Inhibition studies with structural analogs revealed a less specific biotin uptake mechanism in C6 than in NB2a cells. Biocytin normalized deficient biotin-dependent propionyl-CoA carboxylase activity within 4 h in biotin-deficient NB2a cells whereas in C6 cells reactivation was <20% thereby confirming that biocytin is only poorly transported into C6 cells. Specific biocytin uptake into NB2a cells is to our knowledge the first demonstration of a carrier-mediated transport mechanism for this compound. Neuronal biocytin uptake might contribute to the pathogenesis of biotinidase deficiency where biocytin is present in elevated levels.
Authors:
Barbara Baur; Terttu Suormala; E Regula Baumgartner
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Brain research     Volume:  925     ISSN:  0006-8993     ISO Abbreviation:  Brain Res.     Publication Date:  2002 Jan 
Date Detail:
Created Date:  2002-01-16     Completed Date:  2002-03-18     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0045503     Medline TA:  Brain Res     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  111-21     Citation Subset:  IM    
Affiliation:
Metabolic Unit, University Children's Hospital, 4005 Basel, Switzerland.
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MeSH Terms
Descriptor/Qualifier:
Animals
Astrocytoma / metabolism*
Binding, Competitive / physiology
Biological Transport / drug effects,  physiology
Biotin / analogs & derivatives*,  metabolism,  pharmacokinetics*,  pharmacology
Carbon-Carbon Ligases / metabolism
Cell Line, Transformed
Choline / metabolism
Dose-Response Relationship, Drug
Enzyme Activation / drug effects
Lysine / analogs & derivatives*,  pharmacokinetics*
Mice
Neuroblastoma / metabolism*
Neuroglia / cytology,  drug effects,  metabolism*
Neurons / cytology,  drug effects,  metabolism*
Potassium / metabolism
Rats
Sodium / metabolism
Thioctic Acid / pharmacology
Chemical
Reg. No./Substance:
15720-25-9/desthiobiotin; 56-87-1/Lysine; 576-19-2/biocytin; 58-85-5/Biotin; 62-46-4/Thioctic Acid; 62-49-7/Choline; 7440-09-7/Potassium; 7440-23-5/Sodium; EC 6.4.-/Carbon-Carbon Ligases; EC 6.4.1.3/propionyl CoA carboxylase (ATP-hydrolyzing)

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


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