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Updated: Aug 9, 2026

Measuring Plasma Membrane Protein Endocytic Rates by Reversible Biotinylation
Published on: December 24, 2009
Uptake and metabolism of biotin by human peripheral blood mononuclear cells
1Department of Pediatrics (Gastroenterology), University of Arkansas for Medical Sciences and the Arkansas Children's Hospital Research Institute, Little Rock, Arkansas 72202, USA.
Abstract:
We studied the uptake of biotin into human peripheral blood mononuclear cells (PBMC) using [3H]biotin and studied the catabolism of biotin in PBMC using [14C]biotin. Over 30 min, [3H]biotin uptake was greater at 37 degrees C than at 25 degrees C (KT = 2.6 +/- 0.4 nM, maximal velocity = 2.9 +/- 0.2 fmol . 10(6) cells-1 . 30 min-1). Ouabain reduced [3H]biotin uptake to 65% of control values, suggesting that biotin uptake is Na-K-ATPase dependent. Unlabeled biotin and biotin analogs reduced the uptake of [3H]biotin to 22-70% of control values, suggesting the presence of a competition for a structurally specific biotin transporter. When endocytosis by PBMC was stimulated by various acyl glycerols, [3H]biotin uptake was 40-73% of control values; these data are consistent with the hypothesis that stimulated endocytosis reduces biotin transporter density on the cell surface. During a 168-h incubation, PBMC did not catabolize [14C]biotin.
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