Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK310 and Protein G B1.

IF 1.5 4区 工程技术 Q3 ENGINEERING, ELECTRICAL & ELECTRONIC
J E Baio, T Weidner, N T Samuel, Keith McCrea, Loren Baugh, Patrick S Stayton, David G Castner
{"title":"Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK3<sub>10</sub> and Protein G B1.","authors":"J E Baio,&nbsp;T Weidner,&nbsp;N T Samuel,&nbsp;Keith McCrea,&nbsp;Loren Baugh,&nbsp;Patrick S Stayton,&nbsp;David G Castner","doi":"10.1116/1.3456176","DOIUrl":null,"url":null,"abstract":"<p><p>The ability to orient biologically active proteins on surfaces is a major challenge in the design, construction, and successful deployment of many medical technologies. As methods to orient biomolecules are developed, it is also essential to develop techniques that can an accurately determine the orientation and structure of these materials. In this study, two model protein and peptide systems are presented to highlight the strengths of three surface analysis techniques for characterizing protein films: time-of-flight secondary ion mass spectrometry (ToF-SIMS), sum-frequency generation (SFG) vibrational spectroscopy, and near-edge x-ray absorption fine structure (NEXAFS) spectroscopy. First, the orientation of Protein G B1, a rigid 6 kDa domain covalently attached to a maleimide-functionalized self-assembled monolayer, was examined using ToF-SIMS. Although the thickness of the Protein G layer was similar to the ToF-SIMS sampling depth, orientation of Protein G was successfully determined by analyzing the C<sub>2</sub>H<sub>5</sub>S<sup>+</sup> intensity, a secondary ion derived from a methionine residue located at one end of the protein. Next, the secondary structure of a 13-mer leucine-lysine peptide (LK3<sub>10</sub>) adsorbed onto hydrophilic quartz and hydrophobic fluorocarbon surfaces was examined. SFG spectra indicated that the peptide's lysine side chains were ordered on the quartz surface, while the peptide's leucine side chains were ordered on the fluorocarbon surface. NEXAFS results provided complementary information about the structure of the LK3<sub>10</sub> film and the orientations of amide bonds within the LK3<sub>10</sub> peptide.</p>","PeriodicalId":38110,"journal":{"name":"Journal of Vacuum Science and Technology B:Nanotechnology and Microelectronics","volume":null,"pages":null},"PeriodicalIF":1.5000,"publicationDate":"2010-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1116/1.3456176","citationCount":"24","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Vacuum Science and Technology B:Nanotechnology and Microelectronics","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1116/1.3456176","RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"ENGINEERING, ELECTRICAL & ELECTRONIC","Score":null,"Total":0}
引用次数: 24

Abstract

The ability to orient biologically active proteins on surfaces is a major challenge in the design, construction, and successful deployment of many medical technologies. As methods to orient biomolecules are developed, it is also essential to develop techniques that can an accurately determine the orientation and structure of these materials. In this study, two model protein and peptide systems are presented to highlight the strengths of three surface analysis techniques for characterizing protein films: time-of-flight secondary ion mass spectrometry (ToF-SIMS), sum-frequency generation (SFG) vibrational spectroscopy, and near-edge x-ray absorption fine structure (NEXAFS) spectroscopy. First, the orientation of Protein G B1, a rigid 6 kDa domain covalently attached to a maleimide-functionalized self-assembled monolayer, was examined using ToF-SIMS. Although the thickness of the Protein G layer was similar to the ToF-SIMS sampling depth, orientation of Protein G was successfully determined by analyzing the C2H5S+ intensity, a secondary ion derived from a methionine residue located at one end of the protein. Next, the secondary structure of a 13-mer leucine-lysine peptide (LK310) adsorbed onto hydrophilic quartz and hydrophobic fluorocarbon surfaces was examined. SFG spectra indicated that the peptide's lysine side chains were ordered on the quartz surface, while the peptide's leucine side chains were ordered on the fluorocarbon surface. NEXAFS results provided complementary information about the structure of the LK310 film and the orientations of amide bonds within the LK310 peptide.

Abstract Image

Abstract Image

Abstract Image

多肽和蛋白取向的多技术表征:LK310和蛋白gb1。
在许多医疗技术的设计、构建和成功部署中,在表面定向生物活性蛋白质的能力是一个重大挑战。随着生物分子定向方法的发展,开发能够准确确定这些材料的定向和结构的技术也至关重要。在这项研究中,提出了两个模型蛋白质和肽系统,以突出表征蛋白质膜的三种表面分析技术的优势:飞行时间二次离子质谱(ToF-SIMS)、和频生成(SFG)振动光谱和近边缘x射线吸收精细结构(NEXAFS)光谱。首先,使用ToF-SIMS检查蛋白G B1的取向,蛋白G B1是共价连接到马来酰亚胺官能化的自组装单层上的刚性6kDa结构域。尽管G蛋白层的厚度与ToF-SIMS采样深度相似,但通过分析C2H5S+强度(源自位于蛋白质一端的甲硫氨酸残基的二次离子)成功确定了G蛋白的取向。接下来,研究吸附在亲水性石英和疏水性氟碳化合物表面上的13聚亮氨酸赖氨酸肽(LK310)的二级结构。SFG光谱表明,肽的赖氨酸侧链在石英表面有序,而肽的亮氨酸侧链在氟碳化合物表面有序。NEXAFS结果提供了关于LK310膜的结构和LK310肽内酰胺键的取向的补充信息。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
CiteScore
2.70
自引率
0.00%
发文量
146
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信