Temperature profiling of polypeptides in reversed-phase liquid chromatography -: I.: Monitoring of dimerization and unfolding of amphipathic α-helical peptides
Temperature profiling of polypeptides in reversed-phase liquid chromatography -: I.: Monitoring of dimerization and unfolding of amphipathic α-helical peptides
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DOI：
10.1016/s0021-9673(03)00621-6
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发表时间：
2003-08-15
影响因子：
4.1
通讯作者：
Hodges, RS
中科院分区：
文献类型：
--
作者：
Mant, CT;Chen, Y;Hodges, RS
本研究旨在通过证明反相液相色谱法（RP-HPLC）能够监测从头设计的合成两亲性α-螺旋肽在不同疏水性的固定相上的二聚化和展开来扩展其实用性。因此，我们比较了温度（5-80 ℃）对包含两亲性α-螺旋结构、具有L-或D-取代的两亲性α-螺旋结构或非两亲性α-螺旋结构的肽混合物的RP-HPLC（C-8或氰基柱）洗脱行为的影响。通过比较保留行为的螺旋肽的肽的可忽略的二级结构（无规卷曲），我们合理化的“温度分布”通过RP-HPLC可以监测肽分子的关联，无论是通过寡聚化或聚集，或监测解折叠的α-螺旋肽与温度升高。我们认为，构象依赖性的反应的肽，在不断变化的温度下的RP-HIPLC肽的一般分析和纯化，但也为从头设计的肽和蛋白质的影响。(C)2003 Elsevier B. V.保留所有权利。
The present study sets out to extend the utility of reversed-phase liquid chromatography (RP-HPLC) by demonstrating its ability to monitor dimerization and unfolding of de novo designed synthetic amphipathic alpha-helical peptides on stationary phases of varying hydrophobicity. Thus, we have compared the effect of temperature (5-80 degreesC) on the RP-HPLC (C-8 or cyano columns) elution behaviour of mixtures of peptides encompassing amphipathic alpha-helical structure, amphipathic alpha-helical structure with L- or D-substitutions or non-amphipathic alpha-helical structure. By comparing the retention behaviour of the helical peptides to a peptide of negligible secondary structure (a random coil), we rationalize that "temperature profiling" by RP-HPLC can monitor association of peptide molecules, either through oligomerization or aggregation, or monitor unfolding of a-helical peptides with increasing temperature. We believe that the conformation-dependent response of peptides to RP-HIPLC under changing temperature has implications both for general analysis and purification of peptides but also for the de novo design of peptides and proteins. (C) 2003 Elsevier B.V. All rights reserved.