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Phage Library (phage + library)

Distribution by Scientific Domains

Medical Sciences	42%
Chemistry	42%

Selected Abstracts

Phage display as a novel screening tool for primary toxicological targets

ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 2 2010
Bieke Van Dorst
Abstract In the present study the use of phage display as a screening tool to determine primary toxicological targets was investigated. These primary toxicological targets are the targets in the cell with which a chemical compound initially interacts and that are responsible for consecutive (toxic) effects. Nickel was used as model compound for the present study. By selection of Ni-binding peptides out of a 12-mer peptide phage library, it was possible to identify primary toxicological targets of Ni (and other metals). The selected Ni-binding peptides showed similarities to important primary toxicological targets of Ni, such as the hydrogenase nickel incorporation protein (hypB) and the Mg/Ni/Co transporter (corA). This shows that phage display, which is already widely used in other research fields, also has potential in ecotoxicology, as a novel screening tool with which to determine primary toxicological targets of chemical compounds. Environ. Toxicol. Chem. 2010;29:250,255. © 2009 SETAC [source]

Redox-regulated affinity of the third PDZ domain in the phosphotyrosine phosphatase PTP-BL for cysteine-containing target peptides

FEBS JOURNAL, Issue 13 2005
Lieke C. J. Van Den Berk
PDZ domains are protein,protein interaction modules that are crucial for the assembly of structural and signalling complexes. They specifically bind to short C-terminal peptides and occasionally to internal sequences that structurally resemble such peptide termini. The binding of PDZ domains is dominated by the residues at the P0 and P,2 position within these C-terminal targets, but other residues are also important in determining specificity. In this study, we analysed the binding specificity of the third PDZ domain of protein tyrosine phosphatase BAS-like (PTP-BL) using a C-terminal combinatorial peptide phage library. Binding of PDZ3 to C-termini is preferentially governed by two cysteine residues at the P,1 and P,4 position and a valine residue at the P0 position. Interestingly, we found that this binding is lost upon addition of the reducing agent dithiothrietol, indicating that the interaction is disulfide-bridge-dependent. Site-directed mutagenesis of the single cysteine residue in PDZ3 revealed that this bridge formation does not occur intermolecularly, between peptide and PDZ3 domain, but rather is intramolecular. These data point to a preference of PTP-BL PDZ3 for cyclic C-terminal targets, which may suggest a redox state-sensing role at the cell cortex. [source]

Development of recombinant inhibitors specific to human kallikrein 2 using phage-display selected substrates

FEBS JOURNAL, Issue 3 2004
Sylvain M. Cloutier
The reactive site loop of serpins undoubtedly defines in part their ability to inhibit a particular enzyme. Exchanges in the reactive loop of serpins might reassign the targets and modify the serpin,protease interaction kinetics. Based on this concept, we have developed a procedure to change the specificity of known serpins. First, reactive loops are very good substrates for the target enzymes. Therefore, we have used the phage-display technology to select from a pentapeptide phage library the best substrates for the human prostate kallikrein hK2 [Cloutier, S.M., Chagas, J.R., Mach, J.P., Gygi, C.M., Leisinger, H.J. & Deperthes, D. (2002) Eur. J. Biochem. 269, 2747,2754]. Selected substrates were then transplanted into the reactive site loop of ,1-antichymotrypsin to generate new variants of this serpin, able to inhibit the serine protease. Thus, we have developed some highly specific ,1-antichymotrypsin variants toward human kallikrein 2 which also show high reactivity. These inhibitors might be useful to help elucidate the importance of hK2 in prostate cancer progression. [source]

SEREX identification of new tumour-associated antigens in cutaneous T-cell lymphoma

BRITISH JOURNAL OF DERMATOLOGY, Issue 2 2004
T.B. Hartmann
Summary Background Cutaneous T-cell lymphoma (CTCL) is a clonal lymphoproliferative disorder of mainly CD4+ T cells, with primary manifestation in the skin. Objectives To detect new CTCL-associated antigens for immunological therapies and to define their specificity in terms of RNA expression and seroreactivity. Methods A newly constructed CTCL cDNA phage library was screened and cross-reactivities against the detected clones were tested using 15 mycosis fungoides and six Sézary syndrome sera. The mRNA expression of the identified genes was analysed by reverse transcription,polymerase chain reaction (RT,PCR) using 22 tumour tissues, nine cell lines and up to 29 different types of normal tissue. Results We identified nine different tumour antigens (HD-CL-01 to HD-CL-09) of which seven clones had high homology to genes with known functions. Several of these genes had previously been associated with cancer, namely inositol 1,4,5-triphosphate 5-phosphatase, vimentin, aldose reductase and elongation factor-1,. Variations in the deduced protein sequences were observed in three cases, mostly due to variations in protein length. The individual clones were recognized by up to 56% of patients' sera, while control sera were negative except in one case. Using RT,PCR, we found a frequent expression of these new tumour antigens in tumour specimens (26,100%). In contrast to humoral specificity, specific mRNA was also detected in selected normal tissues (29,89%). Conclusions SEREX (serological identification of antigens by recombinant expression cloning) identified multiple tumour-associated antigens in CTCL. The serological specificity and the high percentage of reactive sera of CTCL patients against several clones suggest these genes as potential targets for diagnostic and prognostic purposes. [source]

Identifying Natural Product Biosynthetic Genes from a Soil Metagenome by Using T7 Phage Selection

CHEMBIOCHEM, Issue 16 2009
Keya Zhang
Doing the dirty work: Metagenomic DNA was directly isolated from soil bacteria to construct a T7 phage library. Iterative phage selection by protein modification catalyzed by phosphopantetheinyl transferase enriched putative acyl carrier protein and peptidyl carrier protein genes that are associated with natural product biosynthetic enzymes. [source]

Identification of peptides specific for antibodies in vitiligo using a phage library

CLINICAL & EXPERIMENTAL DERMATOLOGY, Issue 6 2005
Z. Jadali
Summary Patients with vitiligo produce specific autoantibodies that can be detected in their sera. These antibodies are believed to play a role in the pathogenesis of this disease. A random peptide library displayed on phage is a technique that can be used to identify the epitopes that react with monoclonal and polyclonal antibodies. We used this technique to identify the epitopes that react specifically with the vitiligo autoantibodies. By screening the random peptide phage library and using ELISA, two clones that showed a higher frequency of reactivity with the antibodies in the sera of patients with vitiligo were identified. The peptides do not show any similarity with the autoantigens so far implicated in vitiligo, indicating that these epitopes may mimic conformational epitopes in proteins. [source]