Serum levels of thioredoxin/adult T cell leukemia derived factor (TRX/ADF) in asthmatic patients

Yamada, Yoshiyuki; Nakamura, Hajime; Sannohe, Satoshi; Saito, Norihiro; Cui, Chang-Hao; Adachi, Tomohiko; Kayaba, Hiroyuki; Yodoi, Junji; Chihara, Junichi

doi:10.1016/s0091-6749(02)81164-2

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“…Intracellular thiols play essential roles in redox regulation of cell growth, apoptosis inhibition, DNA synthesis, and angiogenesis. − The major thiols of the redox system are glutathione (GSH) and thioredoxin (Trx). , Trx is primarily involved in the reducing process of protein disulfides such as ribonucleotide reductase, methionine sulfoxide reductases, and peroxiredoxins . Furthermore, the overexpression of Trx in intracellular and extracellular compartments has been implicated in several types of cancers, cardiovascular diseases, etc. − In addition, mitochondrial Trx is essential in scavenging oxidatively damaged mitochondrial proteins, regulating mitochondrial permeability transition, − and participating against apoptosis. − Moreover, inappropriate levels of mitochondrial Trx are directly associated with mitochondria dysfunctions, resulting in increased reactive oxygen species, higher apoptosis rate, and lower growth. ,− …”

Section: Introductionmentioning

confidence: 99%

Mitochondrial Thioredoxin-Responding Off–On Fluorescent Probe

et al. 2012

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We synthesized a new probe, Mito-Naph, to visualize mitochondrial thioredoxin (Trx) activity in cells. A fluorescence off-on change is induced by disulfide cleavage of the probe, resulting from a reaction with Trx and subsequent intramolecular cyclization by the released thiolate to give a fluorescent product. By measuring the fluorescence at 540 nm, Trx activity can be detected at nanomolar concentrations (down to 50 nM) well below its physiological levels. The in vitro and in vivo Trx preference of Mito-Naph was demonstrated by fluorometric and confocal microscopic experiments. In vitro kinetic analysis of the disulfide bond cleavage revealed that the second-order rate constant for Trx is (4.04 ± 0.26) × 10(3) (M s)(-1), approximately 5000 times faster than that for GSH. The inhibition experiments involving PX-12, a selective inhibitor of Trx, also revealed that the emission from Mito-Naph significantly decreased in PX-12 dose-dependent manners, both in living cells and in cellular protein extracts. The Trx preference was further supported by an observation that the fluorescence intensity of rat liver extract was decreased according to the Trx depletion by immunoprecipitation. On the basis of these results, it is concluded that Mito-Naph preferentially reacts with Trx, compared with other biological thiols containing amino acids in vitro and in vivo.

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