A solution NMR investigation into the early events of amelogenin nanosphere self-assembly initiated with sodium chloride or calcium chloride.
A solution NMR investigation into the early events of amelogenin nanosphere self-assembly initiated with sodium chloride or calcium chloride.
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DOI：
10.1021/bi8018288
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发表时间：
2008-12-16
期刊：
影响因子：
2.9
通讯作者：
Shaw, Wendy J.
中科院分区：
文献类型：
--
作者：
Buchko, Garry W.;Tarasevich, Barbara J.;Bekhazi, Jacky;Snead, Malcolm L.;Shaw, Wendy J.
使用溶液状态的NMR光谱，新的见解的早期事件的釉原蛋白超分子自组装已被确定使用氯化钠和氯化钙触发协会。二维1H-15 N HSQC光谱记录为15 N-和13 C-标记的鼠釉原蛋白作为增加NaCl和CaCl 2浓度的函数，开始于pH 3.0的2%乙酸溶液条件，其中釉原蛋白是单体。残留物的具体变化，表现为在1H-15 N HSQC交叉峰的强度和消失的分子动力学，观察到与添加任何盐的蛋白质。随着NaCl浓度的增加，T21和R31之间的N-末端附近的残基首先受到影响，这表明这些残基可能启动釉原蛋白二聚化，纳米球组装的第一步。在较高的NaCl浓度下，更多的N-末端附近的残基（Y12-I51）受到影响，并且随着NaCl的进一步添加，C-末端附近的残基（L141-T171）开始显示出类似的分子动力学变化。随着氯化钙浓度的增加，观察到一个类似的逐步变化的分子动力学基本上涉及相同的一组釉原蛋白残基。随着任一盐浓度的增加，观察到估计的总体旋转相关时间（τc）的伴随增加，与组装一致。在类似条件下用动态光散射研究建立了自组装成二聚体或三聚体，所述研究显示最小物质的直径从不存在盐的情况下的4.1 nm增加到存在盐的情况下的约10 nm。这些结果表明一个可能的逐步相互作用机制，从N-末端开始，然后是C-末端，导致釉原蛋白纳米球组装。
Using solution-state NMR spectroscopy, new insights into the early events governing amelogenin supramolecular self-assembly have been identified using sodium chloride and calcium chloride to trigger the association. Two-dimensional 1H–15N HSQC spectra were recorded for 15N- and 13C-labeled murine amelogenin as a function of increasing NaCl and CaCl2 concentration beginning with solution conditions of 2% acetic acid at pH 3.0, where amelogenin was monomeric. Residue specific changes in molecular dynamics, manifested by the reduction in intensity and disappearance of 1H–15N HSQC cross-peaks, were observed with the addition of either salt to the protein. With increasing NaCl concentrations, residues between T21 and R31 near the N-terminus were affected first, suggesting that these residues may initiate amelogenin dimerization, the first step in nanosphere assembly. At higher NaCl concentrations, more residues near the N-terminus (Y12–I51) were affected, and with further additions of NaCl, residues near the C-terminus (L141–T171) began to show a similar change in molecular dynamics. With increasing CaCl2 concentrations, a similar stepwise change in molecular dynamics involving essentially the same set of amelogenin residues was observed. As the concentration of either salt was increased, a concomitant increase in the estimated overall rotational correlation time (τc) was observed, consistent with assembly. Self-assembly into a dimer or trimer was established with dynamic light scattering studies under similar conditions that showed an increase in diameter of the smallest species from 4.1 nm in the absence of salt to ~10 nm in the presence of salt. These results suggest a possible stepwise interaction mechanism, starting with the N-terminus and followed by the C-terminus, leading to amelogenin nanosphere assembly.
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影响因子：
0.9
作者：
Buchko, Garry W.;Bekhazi, Jacky;Shaw, Wendy J.
通讯作者：
Shaw, Wendy J.
影响因子：
2.7
作者：
GOTO, Y;KOGURE, E;AOBA, T
通讯作者：
AOBA, T
影响因子：
2.9
作者：
Mulder, FAA;Hon, B;Kay, LE
通讯作者：
Kay, LE
DOI：
10.1073/pnas.78.6.3824
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
HOPP, TP;WOODS, KR
通讯作者：
WOODS, KR
影响因子：
3
作者：
FINCHAM, AG;MORADIANOLDAK, J;SLAVKIN, HC
通讯作者：
SLAVKIN, HC