Protein-bound NAD(P)H Lifetime is Sensitive to Multiple Fates of Glucose Carbon.
Protein-bound NAD(P)H Lifetime is Sensitive to Multiple Fates of Glucose Carbon.
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DOI：
10.1038/s41598-018-23691-x
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发表时间：
2018-04-03
影响因子：
4.6
通讯作者：
Skala MC
中科院分区：
文献类型：
--
作者：
Sharick JT;Favreau PF;Gillette AA;Sdao SM;Merrins MJ;Skala MC
虽然 NAD(P)H 荧光寿命成像 (FLIM) 可以检测 TCA 循环和电子传递链 (ETC) 的通量变化，但尚不清楚 NAD(P)H FLIM 是否对葡萄糖的其他潜在命运敏感。葡萄糖碳可以通过戊糖磷酸途径（通过葡萄糖 6-磷酸脱氢酶，G6PDH）、乳酸生成（通过乳酸脱氢酶，LDH）和从 TCA 循环中排斥碳（通过丙酮酸脱氢酶激酶，PDK）从线粒体转移，所有这些都可以在癌细胞中上调。在这里，我们证明多光子 NAD(P)H FLIM 可用于定量溶液中重组 LDH 和苹果酸脱氢酶 (MDH) 的相对浓度。在多种上皮细胞系中，NAD(P)H FLIM 对 LDH 和 PDK 的抑制以及被迫使用丙酮酸与乳酸作为燃料来源的细胞中 LDH 的方向性也敏感。在 FLIM 可测量的参数中，只有蛋白质结合的 NAD(P)H (τ2) 的寿命对这些变化敏感，与光学氧化还原比、平均 NAD(P)H 寿命、游离 NAD(P)H 寿命或游离和蛋白质结合的 NAD(P)H 的相对量相反。 NAD(P)H τ2 提供了非侵入性量化碳从 TCA 循环/ETC 转移的能力，这可能支持耐药机制。
While NAD(P)H fluorescence lifetime imaging (FLIM) can detect changes in flux through the TCA cycle and electron transport chain (ETC), it remains unclear whether NAD(P)H FLIM is sensitive to other potential fates of glucose. Glucose carbon can be diverted from mitochondria by the pentose phosphate pathway (via glucose 6-phosphate dehydrogenase, G6PDH), lactate production (via lactate dehydrogenase, LDH), and rejection of carbon from the TCA cycle (via pyruvate dehydrogenase kinase, PDK), all of which can be upregulated in cancer cells. Here, we demonstrate that multiphoton NAD(P)H FLIM can be used to quantify the relative concentrations of recombinant LDH and malate dehydrogenase (MDH) in solution. In multiple epithelial cell lines, NAD(P)H FLIM was also sensitive to inhibition of LDH and PDK, as well as the directionality of LDH in cells forced to use pyruvate versus lactate as fuel sources. Among the parameters measurable by FLIM, only the lifetime of protein-bound NAD(P)H (τ2) was sensitive to these changes, in contrast to the optical redox ratio, mean NAD(P)H lifetime, free NAD(P)H lifetime, or the relative amount of free and protein-bound NAD(P)H. NAD(P)H τ2 offers the ability to non-invasively quantify diversions of carbon away from the TCA cycle/ETC, which may support mechanisms of drug resistance.
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影响因子：
2.2
作者：
Heikal AA
通讯作者：
Heikal AA
DOI：
10.1073/pnas.0611662104
发表时间：
2007-05-29
影响因子：
11.1
作者：
Cairns, Rob A.;Papandreou, Ioanna;Denko, Nicholas C.
通讯作者：
Denko, Nicholas C.
在组织病理学切片中内源性荧光团的荧光寿命成像揭示了乳腺癌原位癌中正常和肿瘤上皮之间的差异。
影响因子：
2.6
作者：
Conklin MW;Provenzano PP;Eliceiri KW;Sullivan R;Keely PJ
通讯作者：
Keely PJ
影响因子：
3.4
作者：
Ma, Ning;Digman, Michelle A.;Gratton, Enrico
通讯作者：
Gratton, Enrico
影响因子：
3.6
作者：
Dimitrow, Enrico;Riemann, Iris;Kaatz, Martin
通讯作者：
Kaatz, Martin