Biocytin and biotin uptake into NB2a neuroblastoma and C6 astrocytoma cells.

Abstract

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
• Journal: Brain research (Brain Res) Vol. 925 Issue 2 Pg. 111-21 (Jan 25 2002) ISSN: 0006-8993 [Print] Netherlands
• PMID: 11792359 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Biotin
• desthiobiotin
• Thioctic Acid
• Sodium
• Carbon-Carbon Ligases
• propionyl CoA carboxylase (ATP-hydrolyzing)
• biocytin
• Lysine
• Choline
• Potassium

Topics

• 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)