Molecular evolution of peptides by yeast surface display technology

Linciano, Sara; Pluda, Stefano; Bacchin, Arianna; Angelini, Alessandro

doi:10.1039/c9md00252a

Cited by 42 publications

(32 citation statements)

References 89 publications

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“…Versatility follows from the huge number of variations available: if one considers the 20 standard (“L”) natural amino acids (a set that can be expanded by considering D isomers and a variety of unnatural amino acids), one can theoretically build 20 N different sequences, where N is the number of monomeric units. Exploiting this variability and selecting the useful sequences is achieved by screening large random libraries selecting effective peptides thanks to phage [ 218 , 219 ], yeast [ 220 ], bacterial [ 221 ] and other forms of display/biopanning technology. Once an effective sequence is identified, it can be readily produced in the lab by the convenient, generally standard methods of solid phase peptide synthesis.…”

Section: Modulation Of Cancer Cell Fate By Peptides Targeting Mitochondrial Ion Channel Interactionsmentioning

confidence: 99%

Targeting mitochondrial ion channels for cancer therapy

2021

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Pharmacological targeting of mitochondrial ion channels is emerging as a promising approach to eliminate cancer cells; as most of these channels are differentially expressed and/or regulated in cancer cells in comparison to healthy ones, this strategy may selectively eliminate the former. Perturbation of ion fluxes across the outer and inner membranes is linked to alterations of redox state, membrane potential and bioenergetic efficiency. This leads to indirect modulation of oxidative phosphorylation, which is/may be fundamental for both cancer and cancer stem cell survival. Furthermore, given the crucial contribution of mitochondria to intrinsic apoptosis, modulation of their ion channels leading to cytochrome c release may be of great advantage in case of resistance to drugs triggering apoptotic events upstream of the mitochondrial phase. In the present review, we give an overview of the known mitochondrial ion channels and of their modulators capable of killing cancer cells. In addition, we discuss state-of-the-art strategies using mitochondriotropic drugs or peptide-based approaches allowing a more efficient and selective targeting of mitochondrial ion channel-linked events.

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Section: Modulation Of Cancer Cell Fate By Peptides Targeting Mitochondrial Ion Channel Interactionsmentioning

confidence: 99%

Targeting mitochondrial ion channels for cancer therapy

2021

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“…To overcome these limitations, we describe the use of yeast surface display for the construction of chemically cyclized peptide libraries that can be subjected to fluorescence activated cell sorting (FACS) to efficiently isolate the highest affinity library clones. Only a few instances are reported of using yeast display to express cyclic peptides, most of which are limited to disulfide-constrained peptides, like knottins 13 . However, to our knowledge, chemical crosslinkers have not been commonly used for the cyclization of yeast-displayed peptides, despite their widespread use for cyclizing mRNA-and phage-display peptide libraries.…”

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confidence: 99%

Isolation of Chemically Cyclized Peptide Binders Using Yeast Surface Display

Bacon

Blain

Burroughs

et al. 2020

Preprint

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Cyclic peptides with engineered protein-binding activity have gained increasing attention for use in therapeutic and biotechnology applications. We describe the efficient isolation and characterization of cyclic peptide binders from genetically encoded combinatorial libraries using yeast surface display. Here, peptide cyclization is achieved by disuccinimidyl glutarate-mediated crosslinking of amine groups within a linear peptide sequence that is expressed as a yeast cell surface fusion. Using this approach, we first screened a library of cyclic heptapeptides by magnetic selection and fluorescence activated cell sorting (FACS), to isolate binders for a model target (lysozyme) with low micromolar binding affinity (K D ~ 1.2 -3.7 µM). The isolated peptides bound lysozyme selectively, and only when cyclized. Importantly, we showed that yeast surface displayed cyclic peptides could be used to efficiently obtain quantitative estimates of binding affinity, without chemical synthesis of the selected peptides. Subsequently, to demonstrate broader applicability of our approach, we isolated cyclic heptapeptides that bind human interleukin-17 (IL-17) using yeast-displayed IL-17 as a target for magnetic selection, followed by FACS using recombinant IL-17. Molecular docking simulations and follow-up experimental analyses identified a candidate cyclic peptide that binds IL-17 in its receptor binding region with moderate affinity (K D ~ 300 nM). Taken together, our results show that yeast surface display can be used to efficiently isolate and characterize cyclic peptides generated by chemical modification from combinatorial libraries.

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“…Nevertheless, their development demonstrates that peptides represent valid molecular modalities for blocking the activity of ADAM and ADAMTS proteins. Indeed, the advent of novel DNA-encoded chemical libraries ( Neri and Lerner, 2018 ) and superior peptide display technologies ( Linciano et al, 2019 ; Sohrabi et al, 2020 ; Peacock and Suga, 2021 ) will enable the high-throughput screening of large combinatorial libraries, facilitating the discovery of novel potent and selective compounds with improved properties on short timescales ( Sohrabi et al, 2020 ). Integration of these powerful combinatorial approaches with better automation, innovative chemical modification strategies and emerging computational methods will contribute to the development of better peptide-based inhibitors against ADAM and ADAMTS proteins, which have the potential to be used in the clinic in the near future.…”

Section: Discussionmentioning

confidence: 99%

Peptide-Based Inhibitors of ADAM and ADAMTS Metalloproteinases

Pluda

Mazzocato

Angelini

2021

Front. Mol. Biosci.

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ADAM and ADAMTS are two large metalloproteinase families involved in numerous physiological processes, such as shedding of cell-surface protein ectodomains and extra-cellular matrix remodelling. Aberrant expression or dysregulation of ADAMs and ADAMTSs activity has been linked to several pathologies including cancer, inflammatory, neurodegenerative and cardiovascular diseases. Inhibition of ADAM and ADAMTS metalloproteinases have been attempted using various small molecules and protein-based therapeutics, each with their advantages and disadvantages. While most of these molecular formats have already been described in detail elsewhere, this mini review focuses solely on peptide-based inhibitors, an emerging class of therapeutic molecules recently applied against some ADAM and ADAMTS members. We describe both linear and cyclic peptide-based inhibitors which have been developed using different approaches ranging from traditional medicinal chemistry and rational design strategies to novel combinatorial peptide-display technologies.

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Molecular evolution of peptides by yeast surface display technology

Abstract: This review provides a detailed analysis of the diverse genetically encoded peptides that have been evolved by using yeast surface display technology.

Cited by 42 publications

References 89 publications

Targeting mitochondrial ion channels for cancer therapy

Targeting mitochondrial ion channels for cancer therapy

Isolation of Chemically Cyclized Peptide Binders Using Yeast Surface Display

Peptide-Based Inhibitors of ADAM and ADAMTS Metalloproteinases

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