Design and synthesis of novel cyclic RGD-containing peptides as highly potent and selective integrin .alpha.IIb.beta.3 antagonists

Cheng, Soan; Craig, William S.; Mullen, Daniel G.; Tschopp, Juerg; Dixon, Douglas B.; Pierschbacher, Michael D.

doi:10.1021/jm00027a001

Cited by 153 publications

(117 citation statements)

References 17 publications

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“…The soluble proteins were used to inhibit platelet aggregation. IC 50 s were 0.6 Ϯ 0.2 and 2.1 Ϯ 0.4 M for MalE-RGD and MalE-EETI-CK RGD , respectively, which are close to the IC 50 of 0.33 M obtained for the cyclic CIPRGDYRC (8). No inhibition of platelet aggregation was observed with MalE-EETI-II (data not shown).…”

Section: Vol 183 2001 E Coli K-12 Surface Display Of Ehec Intimin supporting

confidence: 75%

“…Aggregation was stimulated by adding 25 M ADP, and the change in OD 650 was monitored by photometry. Values were expressed as percent aggregation, which represents the percentage of light transmission standardized to fully and not aggregated samples (8). The IC 50 is the concentration of the protein at which the platelet aggregation is inhibited by 50%.…”

Section: Flow-cytometric Analysis and Facsmentioning

confidence: 99%

“…Many integrins, which are heterodimeric proteins with two membrane-spanning subunits, recognize short arginine-glycine-aspartic acid (RGD)-containing amino acid sequences (41). An RGD sequence resides in the cell attachment site of fibrinogen (44), and integrin binding peptides containing an RGD sequence have been identified from random peptide libraries displayed on phage (25) and from chemically synthesized libraries (8) by panning on purified fibrinogen receptors. To investigate whether an RGD-containing peptide displayed on the E. coli cell surface via fusion to intiminЈ can promote binding of the bacteria to the fibrinogen receptor residing on platelets, two versions of intiminЈ-RGD peptide fusions were constructed (Fig.…”

Section: Vol 183 2001 E Coli K-12 Surface Display Of Ehec Intimin mentioning

confidence: 99%

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Display of Passenger Proteins on the Surface ofEscherichia coliK-12 by the EnterohemorrhagicE. coliIntimin EaeA

et al. 2001

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Intimins are members of a family of bacterial adhesins from pathogenic Escherichia coli which specifically interact with diverse eukaryotic cell surface receptors. The EaeA intimin from enterohemorrhagic E. coli O157:H7 contains an N-terminal transporter domain, which resides in the bacterial outer membrane and promotes the translocation of four C-terminally attached passenger domains across the bacterial cell envelope. We investigated whether truncated EaeA intimin lacking two carboxy-terminal domains could be used as a translocator for heterologous passenger proteins. We found that a variant of the trypsin inhibitor Ecballium elaterium trypsin inhibitor II (EETI-II), interleukin 4, and the Bence-Jones protein REI v were displayed on the surface of E. coli K-12 via fusion to truncated intimin. Fusion protein net accumulation in the outer membrane could be regulated over a broad range by varying the cellular amount of suppressor tRNA that is necessary for translational readthrough at an amber codon residing within the truncated eaeA gene. Intimin-mediated adhesion of the bacterial cells to eukaryotic target cells could be mimicked by surface display of a short fibrinogen receptor binding peptide containing an arginine-glycine-aspartic acid sequence motif, which promoted binding of E. coli K-12 to human platelets. Cells displaying a particular epitope sequence fused to truncated intimin could be enriched 200,000-fold by immunofluorescence staining and fluorescence-activated cell sorting in three sorting rounds. These results demonstrate that truncated intimin can be used as an anchor protein that mediates the translocation of various passenger proteins through the cytoplasmic and outer membranes of E. coli and their exposure on the cell surface. Intimin display may prove a useful tool for future protein translocation studies with interesting biological and biotechnological ramifications.In recent years there has been considerable progress in the development of expression systems for the display of heterologous peptides and proteins on the surfaces of bacteria and yeasts (6,16,31). Cells displaying peptides and proteins such as receptors, antibodies, and enzymes are of considerable value for various biotechnological applications, such as bioseparations, vaccine development, and combinatorial library screening. Numerous anchor proteins that mediate the translocation of passenger proteins through the cytoplasmic and outer membranes of Escherichia coli and their exposure on the cell surface have been used. Short peptides (less than approximately 50 amino acids [aa]) were successfully displayed on the cell surface by insertion into surface-exposed loops of fimbrial proteins (24) or outer membrane proteins like LamB (7) or PhoE (2). Larger passenger domains could be presented on the E. coli cell surface by insertion into a surface-exposed domain of the E. coli flagellin FliC (50), by carboxy-terminal fusion to Lpp-OmpA (a hybrid protein consisting of parts of the E. coli lipoprotein Lpp and OmpA protein [9,12]), by usi...

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Section: Vol 183 2001 E Coli K-12 Surface Display Of Ehec Intimin supporting

confidence: 75%

Section: Flow-cytometric Analysis and Facsmentioning

confidence: 99%

Section: Vol 183 2001 E Coli K-12 Surface Display Of Ehec Intimin mentioning

confidence: 99%

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Display of Passenger Proteins on the Surface ofEscherichia coliK-12 by the EnterohemorrhagicE. coliIntimin EaeA

et al. 2001

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“…The a v h 3 integrin is one of the most specific biomarkers that can differentiate newly formed capillaries from their mature counterparts (67). High-affinity a v h 3 -selective ligands, Arg-Gly-Asp (RGD) in its conformationally constrained cyclic form has a higher binding affinity than in its linear form (68). Ternary doxorubicin-loaded poly(ethylene glycol) nanoparticles conjugated to cyclic RGD (69) and paclitaxel-cyclic RGD nanoparticles have recently been reported.…”

Section: Characteristics Of Cell Surface Receptors and Antigensmentioning

confidence: 99%

Nanotechnology in cancer therapeutics: bioconjugated nanoparticles for drug delivery

Sinha

Kim

Nie

et al. 2006

Molecular Cancer Therapeutics

718

437

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Nanotechnology refers to the interactions of cellular and molecular components and engineered materialstypically, clusters of atoms, molecules, and molecular fragments into incredibly small particles-between 1 and 100 nm. Nanometer-sized particles have novel optical, electronic, and structural properties that are not available either in individual molecules or bulk solids. The concept of nanoscale devices has led to the development of biodegradable self-assembled nanoparticles, which are being engineered for the targeted delivery of anticancer drugs and imaging contrast agents. Nanoconstructs such as these should serve as customizable, targeted drug delivery vehicles capable of ferrying large doses of chemotherapeutic agents or therapeutic genes into malignant cells while sparing healthy cells. Such ''smart'' multifunctional nanodevices hold out the possibility of radically changing the practice of oncology, allowing easy detection and then followed by effective targeted therapeutics at the earliest stages of the disease. In this article, we briefly discuss the use of bioconjugated nanoparticles for the delivery and targeting of anticancer drugs.

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“…It has been previously shown that cells adhere to cycloRGD significantly more strongly (10 times) than to the linearized form. 55 The VnRGD peptide was derived from the fulllength vitronectin protein sequence, containing the RGD sequence 56 based on work done previously. 57 To investigate the adhesive properties of each RGD peptide on ASCs, we performed 3 h-long adhesion assays.…”

Section: Clevenger Et Almentioning

confidence: 99%

Vitronectin-Based, Biomimetic Encapsulating Hydrogel Scaffolds Support Adipogenesis of Adipose Stem Cells

Clevenger

Hinman

Rubin

et al. 2016

Tissue Engineering Part A

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Soft tissue defects are relatively common, yet currently used reconstructive treatments have varying success rates, and serious potential complications such as unpredictable volume loss and reabsorption. Human adipose-derived stem cells (ASCs), isolated from liposuction aspirate have great potential for use in soft tissue regeneration, especially when combined with a supportive scaffold. To design scaffolds that promote differentiation of these cells down an adipogenic lineage, we characterized changes in the surrounding extracellular environment during adipogenic differentiation. We found expression changes in both extracellular matrix proteins, including increases in expression of collagen-IV and vitronectin, as well as changes in the integrin expression profile, with an increase in expression of integrins such as aVb5 and a1b1. These integrins are known to specifically interact with vitronectin and collagen-IV, respectively, through binding to an Arg-Gly-Asp (RGD) sequence. When three different short RGD-containing peptides were incorporated into three-dimensional (3D) hydrogel cultures, it was found that an RGD-containing peptide derived from vitronectin provided strong initial attachment, maintained the desired morphology, and created optimal conditions for in vitro 3D adipogenic differentiation of ASCs. These results describe a simple, nontoxic encapsulating scaffold, capable of supporting the survival and desired differentiation of ASCs for the treatment of soft tissue defects.

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Design and synthesis of novel cyclic RGD-containing peptides as highly potent and selective integrin .alpha.IIb.beta.3 antagonists

Cited by 153 publications

References 17 publications

Display of Passenger Proteins on the Surface ofEscherichia coliK-12 by the EnterohemorrhagicE. coliIntimin EaeA

Display of Passenger Proteins on the Surface ofEscherichia coliK-12 by the EnterohemorrhagicE. coliIntimin EaeA

Nanotechnology in cancer therapeutics: bioconjugated nanoparticles for drug delivery

Vitronectin-Based, Biomimetic Encapsulating Hydrogel Scaffolds Support Adipogenesis of Adipose Stem Cells

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