Chromatopanning for the identification of gallium binding peptides

Schönberger, Nora; Braun, Róbert; Matys, Sabine; Lederer, Franziska L.; Lehmann, Falk; Flemming, Katrin; Pollmann, Katrin

doi:10.1016/j.chroma.2019.04.037

Cited by 13 publications

(6 citation statements)

References 39 publications

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“…The clone C3.129 (HTQHIQSDDHLA) was chosen for cysteine scanning. The peptide sequence presented on the clone promised interesting possibilities for interaction with gallium, and in previous studies [17], a high affinity of the clone for an immobilized gallium target could be demonstrated. Nevertheless, only a low biosorption of free gallium ions could be observed in this work.…”

Section: Resultsmentioning

confidence: 99%

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Directed Evolution and Engineering of Gallium-Binding Phage Clones—A Preliminary Study

et al. 2019

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The phage surface display technology is a useful tool to screen and to extend the spectrum of metal-binding protein structures provided by nature. The directed evolution approach allows identifying specific peptide ligands for metals that are less abundant in the biosphere. Such peptides are attractive molecules in resource technology. For example, gallium-binding peptides could be applied to recover gallium from low concentrated industrial wastewater. In this study, we investigated the affinity and selectivity of five bacteriophage clones displaying different gallium-binding peptides towards gallium and arsenic in independent biosorption experiments. The displayed peptides were highly selective towards Ga3+ whereby long linear peptides showed a lower affinity and specificity than those with a more rigid structure. Cysteine scanning was performed to determine the relationship between secondary peptide structure and gallium sorption. By site-directed mutagenesis, the amino acids of a preselected peptide sequence are systematically replaced by cysteines. The resulting disulphide bridge considerably reduces the flexibility of linear peptides. Subsequent biosorption experiments carried out with the mutants obtained from cysteine scanning demonstrated, depending on the position of the cysteines in the peptide, either a considerable increase in the affinity of gallium compared to arsenic or an increase in the affinity for arsenic compared to gallium. This study shows the impressive effect on peptide–target interaction based on peptide structure and amino acid position and composition via the newly established systematic cysteine scanning approach.

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Section: Resultsmentioning

confidence: 99%

“…In an earlier study, we reported the identification of binding motifs of gallium binding bacteriophage clones obtained from a commercial random peptide library. For the respective clones, a maximum of 14-fold better gallium biosorption compared to wild-type phage binding was achieved [17].…”

Section: Introductionmentioning

confidence: 99%

Directed Evolution and Engineering of Gallium-Binding Phage Clones—A Preliminary Study

et al. 2019

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“…The chromatopanning called biopanning procedure described here was modified from Schönberger et al, 2019 [14] and first published by Nian et al, 2010 [15]. Target material were arsenic oxyanions, arsenous acid and arsenous anions (H 3 AsO 3 − , H 2 AsO 4 − , HAsO 4 2− , AsO 4 3− ) of trivalent As(III) and pentavalent As(V) immobilized on a monolithic ion exchange column (CIM ® QA Disk Monolithic Column, BIA Separations, Ajdovščina, Slovenia) in a chromatographic setup using an Äkta avant 25 FPLC system (GE Healthcare, Amersham, UK).…”

Section: Methodsmentioning

confidence: 99%

“…Titration and amplification of phage were performed as described by . The main steps of chromatopanning and subsequent sequencing are described below, for detailed descriptions please refer to Schönberger et al, 2019 [14].…”

Section: Phage Librarymentioning

confidence: 99%

Application of Next Generation Sequencing (NGS) in Phage Displayed Peptide Selection to Support the Identification of Arsenic-Binding Motifs

Braun

Schönberger

Vinke

et al. 2020

Viruses

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Next generation sequencing (NGS) in combination with phage surface display (PSD) are powerful tools in the newly equipped molecular biology toolbox for the identification of specific target binding biomolecules. Application of PSD led to the discovery of manifold ligands in clinical and material research. However, limitations of traditional phage display hinder the identification process. Growth-based library biases and target-unrelated peptides often result in the dominance of parasitic sequences and the collapse of library diversity. This study describes the effective enrichment of specific peptide motifs potentially binding to arsenic as proof-of-concept using the combination of PSD and NGS. Arsenic is an environmental toxin, which is applied in various semiconductors as gallium arsenide and selective recovery of this element is crucial for recycling and remediation. The development of biomolecules as specific arsenic-binding sorbents is a new approach for its recovery. Usage of NGS for all biopanning fractions allowed for evaluation of motif enrichment, in-depth insight into the selection process and the discrimination of biopanning artefacts, e.g., the amplification-induced library-wide reduction in hydrophobic amino acid proportion. Application of bioinformatics tools led to the identification of an SxHS and a carboxy-terminal QxQ motif, which are potentially involved in the binding of arsenic. To the best of our knowledge, this is the first report of PSD combined with NGS of all relevant biopanning fractions.

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“…49) In a recent study, an improvement of this technique was obtained using phage Qβ, generating particles with increased thermal stability in a norovirus detection model. 50) This evidence, together with the explosive increase in smartphone-based biochemical detection systems, [51][52][53][54][55] might point to a future where inexpensive, fast and easy detection of pathogens, assisted by phages, could become a daily occurrence.…”

Section: Detection Of Pathogens In Clinical Settings and Food Industrymentioning

confidence: 99%

Engineered Bacteriophages for Practical Applications

Pizarro-Bäuerle

Ando

2020

Biological & Pharmaceutical Bulletin

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The diversity of advanced genetic engineering techniques that have become available in recent years has enabled a more precise manipulation of genes and genomes. Among these, bacteriophage genomes stand out as an interesting target due to their dependence on a host for replication, which previously complicated their manipulation, and due as well to the many possible fields in which they can be used. In this review, we highlight recent applications for which genetically modified bacteriophages are being employed: as phage therapy in medicine, animal industries and agricultural settings; as a source of new antimicrobials; as biosensors for research, health and environmental purposes; and as genetic engineering tools themselves.

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Chromatopanning for the identification of gallium binding peptides

Cited by 13 publications

References 39 publications

Directed Evolution and Engineering of Gallium-Binding Phage Clones—A Preliminary Study

Directed Evolution and Engineering of Gallium-Binding Phage Clones—A Preliminary Study

Application of Next Generation Sequencing (NGS) in Phage Displayed Peptide Selection to Support the Identification of Arsenic-Binding Motifs

Engineered Bacteriophages for Practical Applications

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