Probing the interface between biomolecules and inorganic materials using yeast surface display and genetic engineering
Probing the interface between biomolecules and inorganic materials using yeast surface display and genetic engineering
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DOI：
10.1016/j.actbio.2004.11.004
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发表时间：
2005-03-01
影响因子：
9.7
通讯作者：
Belcher, AM
中科院分区：
文献类型：
--
作者：
Peelle, BR;Krauland, EM;Belcher, AM
结合在无机材料表面的多肽在仿生材料方面具有广阔的应用前景，但其作用机理目前尚不清楚。本文报道了多肽与半导体硫化镉界面的序列-活性关系，并提出了广泛适用于研究多肽-固体表面相互作用的方法。我们首次使用酵母表面展示技术与人类全套抗体库结合，并将罕见的单链抗体片段鉴定为CDS结合多肽。使用我们的半定量细胞表面结合分析、定点突变分析和基因工程，我们定义了所展示的多肽的短远端区域，对于CDS结合是必要的和充分的。丙氨酸扫描突变与一系列工程多组氨酸多肽的结合揭示了组氨酸数目与结合强度之间的直接关系，这似乎进一步受到精氨酸和碱性残基的调节。通过对可溶性合成肽类似物的竞争研究，确定了最小相互作用强度，表明酵母与CDS结合的抑制在类似2MU的M肽时达到一半最大。然后，我们展示了展示材料特定多肽的细胞形成自我修复生物膜的能力，并区分制造的异质结构表面的材料。此外，我们还展示了所选择的可溶性硫化镉多肽在室温下催化水相合成荧光硫化镉纳米颗粒的潜力。该平台可进一步用于阐明控制界面相互作用的机制，并产生用于医学、生物传感器和高价值无机材料的生物生产的特定材料试剂。(C)2004年Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Although promising for biomimetic materials applications, polypeptides binding inorganic material surfaces and the mechanism of their function have been difficult to characterize. This paper reports sequence-activity relationships of peptides interfacing with semiconductor CdS, and presents methodologies broadly applicable to studying peptide-solid surface interactions. We first employed yeast surface display with a human repertoire antibody library and identified rarely-occurring scFv fragments as CdS-binding polypeptides. Using our semi-quantitative cell-surface binding assay, site-directed mutational analysis, and genetic engineering we defined short distal regions of the displayed polypeptides necessary and sufficient for CdS binding. Alanine scanning mutagenesis in combination with a series of engineered polyhistidine peptides elucidated a direct relationship between histidine number and binding strength, which appeared to be further modulated by arginine and basic residues. The minimum strength of interaction was established by competition studies using soluble synthetic peptide analogs, which showed half-maximal inhibition of yeast binding to CdS at similar to 2 mu M peptide. We then showed the ability of cells displaying material-specific polypeptides to form self-healing biofilms and discriminate between materials of fabricated heterostructure surfaces. Furthermore, we demonstrated the synthetic potential of the selected soluble CdS peptide in mediating aqueous synthesis of fluorescent CdS nanoparticles at room temperature. This platform may be further applied to elucidate mechanisms governing interfacial interactions and to generate material-specific reagents useful in medicine, biosensors, and bioproduction of high value inorganic materials. (c) 2004 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.