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Letarte, Michelle; Voulgaraki, Despina; Hatherley, Deborah; Foster-Cuevas, Mildred; Saunders, N.; Barclay, A. Neil

doi:10.1186/1471-2091-6-2

Cited by 28 publications

(9 citation statements)

References 36 publications

(50 reference statements)

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“…These methods include oriented display around microbeads (Wright et al 2000;Letarte et al 2005) or tags producing dimers (such as Fcfusion proteins), trimers, and, often most potently, pentamers (Holler et al 2000;Voulgaraki et al 2005). No broad assessment for the suitability of any of these techniques to be used in systematic high-throughput screening has been made, since only individual interactions have been reported (Lin et al 2003;Gonzalez et al 2005).…”

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confidence: 99%

Large-scale screening for novel low-affinity extracellular protein interactions

Söllner²,

et al. 2008

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Extracellular protein-protein interactions are essential for both intercellular communication and cohesion within multicellular organisms. Approximately a fifth of human genes encode membrane-tethered or secreted proteins, but they are largely absent from recent large-scale protein interaction datasets, making current interaction networks biased and incomplete. This discrepancy is due to the unsuitability of popular high-throughput methods to detect extracellular interactions because of the biochemical intractability of membrane proteins and their interactions. For example, cell surface proteins contain insoluble hydrophobic transmembrane regions, and their extracellular interactions are often highly transient, having half-lives of less than a second. To detect transient extracellular interactions on a large scale, we developed AVEXIS (avidity-based extracellular interaction screen), a high-throughput assay that overcomes these technical issues and can detect very transient interactions (halflives Յ 0.1 sec) with a low false-positive rate. We used it to systematically screen for receptor-ligand pairs within the zebrafish immunoglobulin superfamily and identified novel ligands for both well-known and orphan receptors. Genes encoding receptor-ligand pairs were often clustered phylogenetically and expressed in the same or adjacent tissues, immediately implying their involvement in similar biological processes. Using AVEXIS, we have determined the first systematic low-affinity extracellular protein interaction network, supported by independent biological data. This technique will now allow large-scale extracellular protein interaction mapping in a broad range of experimental contexts.

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confidence: 99%

Large-scale screening for novel low-affinity extracellular protein interactions

Söllner²,

et al. 2008

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“…Towards this end, in 2005 Barclay's group first evaluated low-affinity ePPIs in the protein microarray format by using CD200-coupled multivalent microbeads to demonstrate the efficient binding to its receptor CD200R immobilized on epoxy-coated glass slides [49,91]. Recently our own group has further validated this approach by screening a library of 1,334 arrayed proteins against 89 immunoglobulin superfamily (IgSF) receptor baits using bait-Fc fusion proteins bound on Protein A coated microbeads (Figs.…”

Section: Glass Slide-based Protein Microarraysmentioning

confidence: 97%

Protein microarrays, biosensors, and cell-based methods for secretome-wide extracellular protein–protein interaction mapping

Gonzalez¹

2012

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Approximately one quarter of all human genes encode proteins that function in the extracellular space or serve to bridge the extracellular and intracellular environments. Physical associations between these secretome proteins serve to regulate a wide range of biological activities and consequently represent important therapeutic targets. Moreover, some extracellular proteins are targeted by pathogens to allow host access or immune evasion. Despite the importance of extracellular protein-protein interactions, our knowledge in this area has remained sparse. Weak affinities and low abundance have often hindered efforts to identify these interactions using traditional methods such as biochemical purification and cDNA library expression cloning. Moreover, current large-scale protein-protein interaction mapping techniques largely under represent extracellular protein-protein interactions. This review highlights emerging biosensor and protein microarray technology, along with more traditional cell-based techniques, that are compatible with secretome-wide screens for extracellular protein-protein interaction discovery. A combination of these approaches will serve to rapidly expand our knowledge of the extracellular protein-protein interactome.

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“…However, the arrays were faster and simpler to use. Letarte et al [74] examined leukocyte membrane-protein interactions between CD200 and hCD200R proteins using a protein microarray. Anti-hCD200R antibodies were immobilized on the glass surface of the microarray to capture the CD200R.…”

Section: Cell Physiology Research and Confirmation Of Transcriptomicsmentioning

confidence: 99%

Protein microchips in biomedicine and biomarker discovery

et al. 2007

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Protein microarray technology is of high recent interest, especially for generating confirmatory and complementary information for transcriptomic studies. In this paper, the advantages, technical limitations, main application fields, and some early results of protein microarrays are reviewed. Today protein microchip technology is mostly available in the form of printed glass slides, bioaffinity surfaces, and tissue microarray (TMA)-based techniques. The advantages of glass slide-based microchips are the simplicity of their application and their relatively low cost. Affinity surface-based protein chip techniques are applicable to minute amounts of starting material (< 1 microg), but interrogation of these chips requires expensive instrumentation, such as mass spectrometers. TMAs are useful for parallel testing of antibody specificities on a broad range of histological specimens in a single slide. Protein microarrays have been successfully implemented for serum tumor marker profiling, cell physiology studies, and mRNA expression study verification. Some of the bottlenecks of the technology are protein instability, problems with nonspecific interactions, and the lack of amplification techniques to generate sufficient amounts of the lower abundance proteins. In spite of the current difficulties, protein microchips are envisioned to be available for routine biomedical and diagnostic applications provided that the ongoing technological developments are successful in improving sensitivity, specificity, and reducing costs.

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Cited by 28 publications

References 36 publications

Large-scale screening for novel low-affinity extracellular protein interactions

Large-scale screening for novel low-affinity extracellular protein interactions

Protein microarrays, biosensors, and cell-based methods for secretome-wide extracellular protein–protein interaction mapping

Protein microchips in biomedicine and biomarker discovery

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