Monitoring the hydrophilicity/hydrophobicity of amino acid side-chains in the non-polar and polar faces of amphipathic α-helices by reversed-phase and hydrophilic interaction/cation-exchange chromatography
Monitoring the hydrophilicity/hydrophobicity of amino acid side-chains in the non-polar and polar faces of amphipathic α-helices by reversed-phase and hydrophilic interaction/cation-exchange chromatography
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DOI：
10.1016/j.chroma.2004.08.137
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发表时间：
2004-10-22
影响因子：
4.1
通讯作者：
Mant, CT
中科院分区：
文献类型：
--
作者：
Hodges, RS;Chen, Y;Mant, CT
准确监测两亲性α-螺旋肽非极性和极性面上氨基酸取代的疏水性/亲水性效应在合理从头设计更有效的抗菌肽等领域是至关重要的。本文报道了用反相高效液相色谱和亲水相互作用/阳离子交换层析的互补分离模式来监测将单一的L或D-氨基酸取代到具有生物活性的26个氨基酸残基的非极性或极性面中心对多肽的表观亲水性/疏水性和两亲性的影响。我们的结果清楚地表明，反相高效液相色谱法和HILIC/CEX最适合分别在非极性和极性面上进行取代的两亲性多肽的拆分。此外，RP-HPLC和HILIC/CEX被证明是亲水性/疏水性变化的很好的监测器，当这些相应的面上发生氨基酸替换时。我们相信，这些互补的高性能模式为合理设计新型两亲性α-螺旋生物活性多肽提供了极好的潜力。(C)2004爱思唯尔B.V.保留所有权利。
The ability to monitor precisely the hydrophobicity/hydrophilicity effects of amino acid substitutions in both the non-polar and polar faces of amphipathic alpha-helical peptides is critical in such areas as the rational de novo design of more effective antimicrobial peptides. The present study reports our initial results of employing the complementary separation modes of reversed-phase high-performance liquid chromatography (RP-HPLC) and hydrophilic interaction/cation-exchange chromatography (HILIC/CEX) to monitor the effect on apparent peptide hydrophilicity/hydrophobicity and amphipathicity of substituting single L- or D-amino acids into the centre of the non-polar or polar faces of a 26-residue biologically active amphipathic a-helical peptide, V-681. Our results clearly show that RP-HPLC and HILIC/CEX are best suited for resolving amphipathic peptides where substitutions are made in the non-polar and polar faces, respectively. Further, RP-HPLC and HILIC/CEX were demonstrated to be excellent monitors of hydrophilicity/hydrophobicity variations where amino acid substitutions were made in these respective faces. We believe these complementary high-performance modes offer excellent potential for rational design of novel amphipathic alpha-helical biologically active peptides. (C) 2004 Elsevier B.V. All rights reserved.