General method for rapid synthesis of multicomponent peptide mixtures

Furka, Árpád; Sebestyén, Ferenc; Asgedom, Mamo; Dibó, Gábor

doi:10.1111/j.1399-3011.1991.tb00765.x

Cited by 1,108 publications

(255 citation statements)

References 19 publications

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“…Glycopeptides can be generated in a library format via a combinatorial approach [7,8]. The most frequently implemented method for the generation of ''one-bead-one-compound" (glyco)peptide libraries [9,10] is the split-and-mix method [11,12]. This method, combined with the ladder synthesis strategy, offers facile synthesis and characterization of thousands of possible ligands that can be used for interaction studies or the development of new therapeutic agents.…”

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confidence: 99%

Assessing the inhibitory potency of galectin ligands identified from combinatorial (glyco)peptide libraries using surface plasmon resonance spectroscopy

Maljaars¹,

André²,

Halkes³

et al. 2008

Analytical Biochemistry

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a b s t r a c tCombinatorial (glyco)peptide libraries offer the possibility to define effective inhibitors of protein (lectin)-glycan interactions. If a (glyco)peptide surpasses the inhibitory potency of the free sugar, then the new peptide-lectin contacts underlying the affinity enhancement may guide further rational drug design. Focusing on the adhesion/growth regulatory human galectins 1 and 3, a screening of three combinatorial solid-phase (glyco)peptide libraries, containing Gal(b1-O)Thr, Gal(b1-S)Cys/Gal(b1-N)Asn, and Lac(b1-O)Thr, with the fluorescently labeled lectins had led to a series of lead compounds. To define the inhibitory potency of a selection of resynthesized (glyco)peptides systematically, a surface plasmon resonance-based inhibition assay with immobilized asialofetuin was set up. (Glyco)Peptides with up to 66-fold potency relative to free lactose as inhibitor were characterized. The presence of lactose in the most effective glycopeptides indicated the presence of affinity-enhancing peptide-lectin contacts. In addition to drug design, they may be helpful for fine-structural analysis of the binding sites.

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confidence: 99%

Assessing the inhibitory potency of galectin ligands identified from combinatorial (glyco)peptide libraries using surface plasmon resonance spectroscopy

Maljaars¹,

André²,

Halkes³

et al. 2008

Analytical Biochemistry

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“…Solid phase synthesis using the "split-and-mix" technique has become a method of choice in preparation of One-Bead-One-Compound (OBOC) libraries [1,2]. The advantage of the OBOC method is that thousands (or even millions) of peptidic compounds can be rapidly synthesized and screened for biological activity.…”

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confidence: 99%

Sensitive electrospray mass spectrometry analysis of one-bead-one-compound peptide libraries labeled by quaternary ammonium salts

et al. 2012

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A rapid and straightforward method for highthroughput analysis of single resin beads from one-beadone-compound combinatorial libraries with high resolution electrospray ionization tandem mass spectrometry (HR ESI-MS/MS) is presented. The application of an efficient method of peptide derivatization by quaternary ammonium salts (QAS) formation increases ionization efficiency and reduces the detection limit, allowing analysis of trace amounts of compounds by ESI-MS. Peptides, synthesized on solid support, contain a new cleavable linker composed of a Peg spacer (9-aza-3,6,12,15-tetraoxa-10-on-heptadecanoic acid), lysine with ε-amino group marked by the N,N,N-triethylglycine salt, and methionine, which makes possible the selective cleavage by cyanogen bromide. Even a small portion of peptides derivatized by QAS cleaved from a single resin bead is sufficient for sequencing by HR ESI-MS/MS experiments. The developed strategy was applied to a small training library of α chymotrypsin substrates. The obtained results confirm the applicability of the proposed method in combinatorial chemistry. Keywords Derivatization of peptides · ESI-MS/MS

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“…The CPLL were synthesized on beads by the splitcouple-recombine method described in earlier works (Furka et al, 1991;Lam et al, 1991;Buettner et al, 1996). Use of the 20 naturally occurring amino acids theoretically resulted in 64 million different ligands, with each ligand fixed to a single bead (Thulasiraman et al, 2005;Boschetti and Righetti, 2008a).…”

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Looking Deep Inside: Detection of Low-Abundance Proteins in Leaf Extracts of Arabidopsis and Phloem Exudates of Pumpkin

et al. 2012

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The field of proteomics suffers from the immense complexity of even small proteomes and the enormous dynamic range of protein concentrations within a given sample. Most protein samples contain a few major proteins, which hamper in-depth proteomic analysis. In the human field, combinatorial hexapeptide ligand libraries (CPLL; such as ProteoMiner) have been used for reduction of the dynamic range of protein concentrations; however, this technique is not established in plant research. In this work, we present the application of CPLL to Arabidopsis (Arabidopsis thaliana) leaf proteins. One-and two-dimensional gel electrophoresis showed a decrease in high-abundance proteins and an enrichment of less abundant proteins in CPLL-treated samples. After optimization of the CPLL protocol, mass spectrometric analyses of leaf extracts led to the identification of 1,192 proteins in control samples and an additional 512 proteins after the application of CPLL. Upon leaf infection with virulent Pseudomonas syringae DC3000, CPLL beads were also used for investigating the bacterial infectome. In total, 312 bacterial proteins could be identified in infected Arabidopsis leaves. Furthermore, phloem exudates of pumpkin (Cucurbita maxima) were analyzed. CPLL prefractionation caused depletion of the major phloem proteins 1 and 2 and improved phloem proteomics, because 67 of 320 identified proteins were detectable only after CPLL treatment. In sum, our results demonstrate that CPLL beads are a time-and cost-effective tool for reducing major proteins, which often interfere with downstream analyses. The concomitant enrichment of less abundant proteins may facilitate a deeper insight into the plant proteome.

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General method for rapid synthesis of multicomponent peptide mixtures

Cited by 1,108 publications

References 19 publications

Assessing the inhibitory potency of galectin ligands identified from combinatorial (glyco)peptide libraries using surface plasmon resonance spectroscopy

Assessing the inhibitory potency of galectin ligands identified from combinatorial (glyco)peptide libraries using surface plasmon resonance spectroscopy

Sensitive electrospray mass spectrometry analysis of one-bead-one-compound peptide libraries labeled by quaternary ammonium salts

Looking Deep Inside: Detection of Low-Abundance Proteins in Leaf Extracts of Arabidopsis and Phloem Exudates of Pumpkin

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