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Measuring picomolar intracellular exchangeable zinc in PC-12 cells using a ratiometric fluorescence biosensor.

Abstract：

:Zinc plays both physiological and pathological roles in biology, making it of increasing interest. To date, intracellular free zinc has been measured in cell types supplemented with or enriched in zinc, such as hippocampal neurons. Here we quantitatively image intracellular exchangeable zinc in an ordinary resting cell culture line (PC-12), using an excitation ratiometric fluorescent biosensor based on carbonic anhydrase (CA). Human CA II has a K d of 4 pM for zinc and suffers no interference from millimolar calcium or magnesium ions. The CA-based biosensor was readily introduced into the cell by a novel approach: fusing a transactivator of transcription (TAT)-derived cell penetrating peptide to the CA molecule and adding it to the cells. Our results indicate that the resting concentration is approximately 5-10 pM in cytoplasm and nucleus. Interestingly, the tetrakis(2-pyridylmethyl)ethylenediamine (TPEN)-Zn complex and TPEN are both apoptogenic for this cell line.

journal_name

ACS Chem Biol

journal_title

ACS chemical biology

authors

Bozym RA,Thompson RB,Stoddard AK,Fierke CA

doi

10.1021/cb500043a

subject

Has Abstract

pub_date

2006-03-17 00:00:00

pages

103-11

issue

2

eissn

1554-8929

issn

1554-8937

journal_volume

1

pub_type

杂志文章

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