Lactic acid and protein interactions: implications for the NMR visibility of lactate in biological systems
Lactic acid and protein interactions: implications for the NMR visibility of lactate in biological systems
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DOI：
10.1016/s0304-4165(98)00154-8
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发表时间：
1999-01-04
影响因子：
3
通讯作者：
Forder, JR
中科院分区：
文献类型：
--
作者：
Chatham, JC;Forder, JR
将牛血清白蛋白（BSA）加入到乳酸和丙氨酸的溶液中，导致乳酸的H-1-NMR共振消失，而丙氨酸则没有。随着温度的升高，乳酸变得越来越核磁共振可见，加热到65℃以上，冷却到25℃后，乳酸结合减少。当BSA浓度为0.2 mM时，在0.2-35 mM的乳酸浓度范围内（斜率0.384 +/- 0.003），NMR可见乳酸与总乳酸之间存在线性关系。60%添加的乳酸在h -1核磁共振光谱中不可见。然而，在0.1 mM BSA溶液中，斜率明显更高，这表明在这些条件下只有25-30%的乳酸盐是NMR不可见的。本研究的结果表明，乳酸在蛋白质溶液中的核磁共振可见性降低是由于非特异性结合，这取决于蛋白质的三级结构。这不仅对体内h -1核磁共振实验的解释，而且对碳-13和碳-14代谢的研究具有重要意义。(C) 1999 Elsevier Science B.V.版权所有
The addition of bovine serum albumin (BSA) to a solution of lactate and alanine resulted in the disappearance of the H-1-NMR resonances from lactate but not alanine. As temperature is increased lactate becomes increasingly NMR visible and after heating above 65 degrees C and cooling to 25 degrees C lactate binding is reduced. With a concentration of 0.2 mM BSA, there was a linear relationship between NMR visible lactate versus total lactate over a range of lactate concentrations of 0.2-35 mM (slope 0.384 +/- 0.003) indicating that approx. 60% of the added lactate is not visible in the H-1-NMR spectrum. With a 0.1 mM BSA solution, however, the slope was markedly higher indicating that under these conditions only 25-30% of the lactate was NMR invisible. The results from this study indicate that decreased NMR visibility of lactate in proteinaceous solutions is due to non-specific binding which is dependent on the tertiary structure of the protein. This has important implications not only for the interpretation of in vivo H-1-NMR experiments but also for C-13, and C-14 studies of metabolism. (C) 1999 Elsevier Science B.V. All rights reserved.