Aqueous Two-Phase Partitioning for Proteomic Monitoring of Cell Surface Biomarkers in Human Peripheral Blood Mononuclear Cells

2006 ◽  
Vol 5 (5) ◽  
pp. 1168-1175 ◽  
Author(s):  
Henrik Everberg ◽  
Ragna Peterson ◽  
Sabina Rak ◽  
Folke Tjerneld ◽  
Cecilia Emanuelsson
Keyword(s):  
Cell Surface ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Two Phase ◽  
Peripheral Blood Mononuclear ◽  
Phase Partitioning ◽  
Blood Mononuclear Cells

Uptake and metabolism of biotin by human peripheral blood mononuclear cells

1998 ◽  
Vol 275 (2) ◽  
pp. C382-C388 ◽  
Author(s):  
Janos Zempleni ◽  
Donald M. Mock
Keyword(s):  
Cell Surface ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Maximal Velocity ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Uptake And Metabolism

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°C than at 25°C ( K T = 2.6 ± 0.4 nM, maximal velocity = 2.9 ± 0.2 fmol ⋅ 106cells−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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