The sodium/iodide symporter (NIS) is primarily responsible for iodide accumulation in the thyroid gland for the synthesis of thyroid hormones; however, it can also transport other lyotropic anions in the thyroid gland and nonthyroid tissues. Some NIS substrates have important physiological or clinical roles, and others are environmental contaminants with health-related consequences. The aim of this study was to assess the utility of a yellow fluorescent protein variant, YFP-H148Q/I152L, as a biosensor to monitor the cellular uptake of NIS substrates, including thiocyanate (SCN-), nitrate (NO3-), chlorate (ClO3-), perchlorate (ClO4-), and perrhenate (ReO4-). The fluorescence of purified YFP-H148Q/I152L was suppressed by anions with an order of potency of ReO4- > ClO4- = I- = SCN- = ClO3- > NO3- ≫ Cl-. Anions also suppressed the fluorescence of YFP-H148Q/I152L expressed in FRTL-5, a thyroid cell line with high NIS expression. Quantitation of intracellular concentrations revealed differences among anions in the affinity and maximal velocity of NIS-mediated uptake as well as in the rate constant for passive efflux. These results suggest that YFP-H148Q/I152L can serve as an intracellular biosensor of NIS-transported anions and may be useful to study the physiology of endogenous anions as well as the health-related consequences of environmental anions.

Intracellular anion fluorescence assay for sodium/iodide symporter substrates / Di Bernardo J.; Iosco C.; Rhoden K.J.. - In: ANALYTICAL BIOCHEMISTRY. - ISSN 0003-2697. - STAMPA. - 415:(2011), pp. 32-38. [10.1016/j.ab.2011.04.017]

Intracellular anion fluorescence assay for sodium/iodide symporter substrates

DI BERNARDO, JULIE;IOSCO, CARMELA;RHODEN, KERRY JANE
2011

Abstract

The sodium/iodide symporter (NIS) is primarily responsible for iodide accumulation in the thyroid gland for the synthesis of thyroid hormones; however, it can also transport other lyotropic anions in the thyroid gland and nonthyroid tissues. Some NIS substrates have important physiological or clinical roles, and others are environmental contaminants with health-related consequences. The aim of this study was to assess the utility of a yellow fluorescent protein variant, YFP-H148Q/I152L, as a biosensor to monitor the cellular uptake of NIS substrates, including thiocyanate (SCN-), nitrate (NO3-), chlorate (ClO3-), perchlorate (ClO4-), and perrhenate (ReO4-). The fluorescence of purified YFP-H148Q/I152L was suppressed by anions with an order of potency of ReO4- > ClO4- = I- = SCN- = ClO3- > NO3- ≫ Cl-. Anions also suppressed the fluorescence of YFP-H148Q/I152L expressed in FRTL-5, a thyroid cell line with high NIS expression. Quantitation of intracellular concentrations revealed differences among anions in the affinity and maximal velocity of NIS-mediated uptake as well as in the rate constant for passive efflux. These results suggest that YFP-H148Q/I152L can serve as an intracellular biosensor of NIS-transported anions and may be useful to study the physiology of endogenous anions as well as the health-related consequences of environmental anions.
2011
Intracellular anion fluorescence assay for sodium/iodide symporter substrates / Di Bernardo J.; Iosco C.; Rhoden K.J.. - In: ANALYTICAL BIOCHEMISTRY. - ISSN 0003-2697. - STAMPA. - 415:(2011), pp. 32-38. [10.1016/j.ab.2011.04.017]
Di Bernardo J.; Iosco C.; Rhoden K.J.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/102755
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