Tight‐binding inhibitory sequences against pp60<sup><i>c−src</i></sup> identified using a random 15‐amino‐acid peptide library

Nishi, Toru; Budde, Raymond J.A.; McMurray, John S.; Obeyesekere, Nihal U.; Safdar, Naueen; Levin, Victor A.; Saya, Hideyuki

doi:10.1016/s0014-5793(96)01329-4

Cited by 41 publications

(20 citation statements)

References 18 publications

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“…A phage-displayed random peptide library expressing pentadecapeptides at the N terminus of pIII coat protein of M13 phage was kindly provided by Dr Hideyuki Saya of Kumamoto University (Nishi et al, 1996). In vivo biopanning was performed by a modi®ed method as described by Pasqualini et al (1997).…”

Section: Methodsmentioning

confidence: 99%

Anti-neovascular therapy using novel peptides homing to angiogenic vessels

et al. 2002

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Cancer chemotherapy targeted to angiogenic vessels is expected to cause indirect tumor regression through the damage of the neovasculature without the induction of drug resistance. To develop a tool for neovasculaturespeci®c drug delivery, we isolated novel peptides homing to angiogenic vessels formed by a dorsal air sac method from a phage-displayed peptide library. Three distinct phage clones that markedly accumulated in murine tumor xenografts presented PRPGAPLAGSWPGTS-, DRWRPALPVVLFPLH-or ASSSYPLIHWRPWARpeptide respectively. After the determination of the epitope sequences of these peptides, we modi®ed liposomes with epitope penta-peptides. Liposome modi®ed with APRPG-peptide showed high accumulation in murine tumor xenografts, and APRPG-modi®ed liposome encapsulating adriamycin eectively suppressed experimental tumor growth. Finally, speci®c binding of APRPG-modi®ed liposome to human umbilical endothelial cells, and that of PRP-containing peptide to angiogenic vessels in human tumors, i.e., islet cell tumor and glioblastoma, were demonstrated. The present study indicates the usefulness of APRPG-peptide as a tool for anti-neovascular therapy, a novel modality of cancer treatment.

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Section: Methodsmentioning

confidence: 99%

Anti-neovascular therapy using novel peptides homing to angiogenic vessels

et al. 2002

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“…The number of primary clones in the 15-mer library is 2.5 ϫ 10 8 (23), and in the case of the 6-mer library it is 2 ϫ 10 8 (22). Approximately 10 10 phage/well in 100 l of TBS buffer were incubated for 60 min at room temperature.…”

Section: Selection Of Peptides From Phage Display Libraries-microtitermentioning

confidence: 99%

Peptide Ligands for the Fibronectin Type II Modules of Matrix Metalloproteinase 2 (MMP-2)

Trexler

Briknarová

Gehrmann

et al. 2003

Journal of Biological Chemistry

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The interaction of matrix metalloproteinase 2 (MMP-2) with gelatin is mediated by three repeats homologous to fibronectin type II (FN2) modules, which are inserted in the catalytic domain in proximity of the active site. We screened a random 15-mer phage display library to identify peptides that interact with the FN2 modules of MMP-2. Interestingly, the selected peptides are not gelatin-like and do not share a common, obvious sequence motif. However, they contain a high proportion of aromatic residues. The interactions of two peptides, WHWRH0RIPLQLAAGR and THSHQWRHHQF-PAPT, with constructs comprising the in-tandem first and second and second and third FN2 modules of MMP-2 (Col-12 and Col-23, respectively) were characterized by NMR. Both peptides interact with Col-12 and Col-23 with apparent association constants in the mM ؊1 range. Peptide binding results in perturbation of signals from residues located in the gelatin-binding pocket and flexible parts of the molecule. Although the former finding suggests that the gelatin-binding site is involved in the contact, the interpretation of the latter is less straightforward and may well reflect both the direct and indirect effects of the interaction.Matrix metalloproteinase 2 (MMP-2, 1 gelatinase A), and the closely related MMP-9 (gelatinase B) are unique among the metalloproteinases in that three gelatin-binding fibronectin type II (FN2) modules (Col-1, Col-2, and Col-3) are inserted in their catalytic domain in the vicinity of the active site (1). The solution conformation of each FN2 repeat from human MMP-2 has been characterized via NMR spectroscopy (2-4). Moreover, the x-ray crystallographic structure of the intact human pro-MMP-2 has been reported (5).In the second FN2 module from each MMP-2 and MMP-9, residues that are important for the interaction with gelatin have been identified via site-directed mutagenesis (6, 7). Additionally, the ligand binding surfaces of all three modules of MMP-2 have been mapped from 1 H and 15 N NMR perturbations induced by (PPG) 6 and the longer chain analog, (PPG) 12 , synthetic peptide mimics of gelatin (2-4). In line with the crystallographic evidence, which shows that the FN2 modules in MMP-2 point away from each other (5), our NMR studies of the interaction between Col domains and (PPG) 6 and (PPG) 12 have shown that consecutive Col modules contain distinct ligand-binding sites in which affinities for these ligands are virtually identical to those of the individual domains (3,4,8).Although the affinity of the MMP-2 Col domains for collagenous ligands appears by now to be well established, less is known regarding the specificity of the interaction. In our previous studies we found that the peptide PIIKFPGDVA, which corresponds to segment 33-42 of the pro-MMP-2, interacts with the three Col domains of MMP-2 in a manner that mimics the interaction with the collagen-like (PPG) 6 and (PPG) 12 peptides (3, 4). Preference for binding to Col-3 was indicated, consistent with the x-ray crystallographic structure of the pro-MMP-2 (5). In ...

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“…Random peptide library f3-15mer (20), containing 2.5 × 10 8 primary clones (GenBank Accession AF246445) with foreign 15-mer peptide displayed on pIII was provided by George P. Smith (Columbia, MO). Kanamycin resistant E. coli K91BluKan (K91BK) host bacteria were also provided by George P. Smith.…”

Section: Methodsmentioning

confidence: 99%

Identification of Peptide Ligands for Targeting to the Blood-Brain Barrier

et al. 2010

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PurposeTransport of drugs to the brain is limited by the blood-brain barrier. New, specific brain endothelium ligands can facilitate brain-specific delivery of drugs.MethodsWe used phage display in an in situ brain perfusion model to screen for new brain endothelium peptide ligands.ResultsTwo phage clones, displaying 15 amino acid-peptides (GLA and GYR) that were selected for brain binding in the mouse model, showed significant binding to human brain endothelium (hCMEC/D3), compared to a random control phage. This binding was not seen for other human endothelial cells (HUVEC). Binding to hCMEC/D3 cells was dose dependent. When phage GLA and GYR were individually perfused through the murine brain, their ability to bind to the brain was 6-fold (GLA) and 5-fold (GYR) higher than the control phage. When compared to lung perfusion, phage showed an 8.5-fold (GYR) and 48-fold (GLA) preference for brain over lung compared to the control.ConclusionsThese results indicate that two new peptide ligands have been identified that may be used for specific targeting of drugs to the blood-brain barrier.

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Tight‐binding inhibitory sequences against pp60^c−src identified using a random 15‐amino‐acid peptide library

Cited by 41 publications

References 18 publications

Anti-neovascular therapy using novel peptides homing to angiogenic vessels

Anti-neovascular therapy using novel peptides homing to angiogenic vessels

Peptide Ligands for the Fibronectin Type II Modules of Matrix Metalloproteinase 2 (MMP-2)

Identification of Peptide Ligands for Targeting to the Blood-Brain Barrier

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Tight‐binding inhibitory sequences against pp60c−src identified using a random 15‐amino‐acid peptide library

Cited by 41 publications

References 18 publications

Anti-neovascular therapy using novel peptides homing to angiogenic vessels

Anti-neovascular therapy using novel peptides homing to angiogenic vessels

Peptide Ligands for the Fibronectin Type II Modules of Matrix Metalloproteinase 2 (MMP-2)

Identification of Peptide Ligands for Targeting to the Blood-Brain Barrier

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Product

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Tight‐binding inhibitory sequences against pp60^c−src identified using a random 15‐amino‐acid peptide library