【目的】表面プラズモン共鳴センサを用いて、タンパク質畳み込み解除に対する2450MHzマイクロ波の影響を調べること。【方法】タンパク質畳み込み解除の経路で起きるタンパク質とシャペロンの結合を表面プラズモン共鳴により計測する方法を用いた。あるレベルより高い温度へのばく露ストレスによりαクリスタリンのシャペロンに強く結合するクエン酸合成酵素を実験対象のタンパク質として用いた。【結果】 マイクロ波加熱の結果を、温水での加温、およびマイクロ波と温水の組み合わせ加温(目標温度は同じ)に結果と比較した。加温中の温度分布を測定した。マイクロ波は、同じ最高温度まで加温されたタンパク質溶液において、通常の熱ストレスに比べ有意に高い程度の畳み込み解除を生じさせることが示された。
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To study the effect of microwaves at 2450 MHz on protein (citrate synthase) unfolding.
The protein was exposed to microwaves only, conventional heat (water bath) only and combinations of heat and microwaves.
Unfolding proteins tend to bind to chaperones on their unfolding pathway and this attachment is monitored by surface plasmon resonance.
ばく露 | パラメータ |
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ばく露1:
2.45 GHz
Modulation type:
pulsed
ばく露時間:
continuous for 10, 15, and 20 s
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|
The protein solution was subjected to microwave exposure, water bath for 5 min at 34 to 82.5 °C, and water bath at 29.5 to 44 °C followed by microwave exposure.
周波数 | 2.45 GHz |
---|---|
タイプ |
|
ばく露時間 | continuous for 10, 15, and 20 s |
Modulation type | pulsed |
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Repetition frequency | 50 Hz |
Additional information |
sharply peaked waveform having a peak to average ratio between 3:1 and 8:1 [Matsumoto et al., 2005] |
ばく露の発生源/構造 |
|
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チャンバの詳細 | A shelf made from high density polyethylene with holes fitting the Eppendorf tubes was located in the microwave oven cavity with a water ballast placed under it in a fixed spot in order to stabilise the field. |
ばく露装置の詳細 | The protein solution in an Eppendorf tube of 80 µl volume was placed in the hole determined to have the maximum temperature rise. |
Additional information | The temperature rise near the end-tip of the Eppendorf tube was measured over a volume of 0.8 mm side length using a fluoroptic probe. |
測定量 | 値 | 種別 | Method | Mass | 備考 |
---|---|---|---|---|---|
SAR | 4.85 kW/kg | mean | - | - | - |
The data showed that microwaves caused a significantly higher degree of unfolding than conventional thermal stress for protein solutions heated to the same maximum temperature. The data support the hypothesis that microwaves have a non-thermal effect on protein conformation that could take the form of a direct interaction of the electromagnetic field with the protein or its water of hydration.
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