Selection of peptides with surface affinity for α-chymotrypsin using a phage display library

Krook, Margareta; Lindbladh, Christer; Birnbaum, Staffan; Naess, H.; Eriksen, Jon Amund; Mosbach, Klaus

doi:10.1016/0021-9673(95)00270-w

Cited by 20 publications

(12 citation statements)

References 20 publications

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“…However, in contrast to the strategy used here, that method favors the isolation of substrates rather than inhibitors 34 . Peptides that bind to chymotrypsin and trypsin recently have been recovered from random phage-displayed libraries, but the isolated peptides also contained identifiable protease substrate sequences 37,38 . The cyclic conformation is critical for the activity of the CTTHWGFTLC peptide, and libraries that express cyclic peptides may be particularly well suited for identifying peptide inhibitors for metalloproteinases.…”

Section: Ctthwgftlc Inhibits Tumor Growth In Mouse Models (A) Mda-mentioning

confidence: 99%

Tumor targeting with a selective gelatinase inhibitor

et al. 1999

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Several lines of evidence suggest that tumor growth, angiogenesis, and metastasis are dependent on matrix metalloproteinase (MMP) activity. However, the lack of inhibitors specific for the type IV collagenase/gelatinase family of MMPs has thus far prevented the selective targeting of MMP-2 (gelatinase A) and MMP-9 (gelatinase B) for therapeutic intervention in cancer. Here, we describe the isolation of specific gelatinase inhibitors from phage display peptide libraries. We show that cyclic peptides containing the sequence HWGF are potent and selective inhibitors of MMP-2 and MMP-9 but not of several other MMP family members. Our prototype synthetic peptide, CTTHWGFTLC, inhibits the migration of human endothelial cells and tumor cells. Moreover, it prevents tumor growth and invasion in animal models and improves survival of mice bearing human tumors. Finally, we show that CTTHWGFTLC-displaying phage specifically target angiogenic blood vessels in vivo. Selective gelatinase inhibitors may prove useful in tumor targeting and anticancer therapies.

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Section: Ctthwgftlc Inhibits Tumor Growth In Mouse Models (A) Mda-mentioning

confidence: 99%

Tumor targeting with a selective gelatinase inhibitor

et al. 1999

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“…If the peptides identified in this study also increase the activity of PSA against natural substrates, they are potentially useful for studying the biological role of PSA in prostate pathology and physiology. Interestingly, none of the peptides identified inhibited enzyme activity, whereas peptides binding to chymotrypsin and MMP-9 identified by phage peptide library screening inhibit their enzyme activity [46,47].…”

Section: Isobm-mabmentioning

confidence: 99%

Identification of novel prostate‐specific antigen‐binding peptides modulating its enzyme activity

Leinonen

Koivunen³

et al. 2000

European Journal of Biochemistry

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Prostate-specific antigen (PSA) is a serine protease with highly prostate-specific expression. Measurement of PSA in serum is widely used for diagnosis and monitoring of prostate cancer. PSA dissolves the seminal gel forming after ejaculation. It has been suggested to mediate invasion and metastasis of prostate cancer but also to exert antiangiogenic activity. We have identified peptides specific for PSA by screening cyclic phage display peptide libraries. PSA-binding peptides were isolated from four different libraries and produced as a fusion protein with glutathione S-transferase (GST). The phage and fusion proteins were shown to bind to PSA specifically as indicated by lack of binding to other serine proteinases. A peptide with four cysteines showed the highest affinity for PSA. Zn 21 , an inhibitor of PSA activity, increased the affinity of the peptides to PSA. The binding specificity was characterized by cross-inhibition using monoclonal anti-PSA antibodies of known epitope specificities. The peptides bound to the same region as mAbs specific for free PSA indicating that they bind close to the active site of the enzyme. The peptides enhanced the enzyme activity of PSA against a chromogenic substrate. These results show that peptides binding to PSA and modulating its enzyme activity can be developed by phage display technique. The peptides have the potential to be used for identification of PSA variants and for imaging and targeting of prostatic tumors.

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“…Selection from phage displayed peptide libraries 5,6 has led to the discovery of peptide mimotopes, which mimic small molecules such as biotin or polysaccharides and nonlinear protein epitopes [7][8][9][10][11] . These peptides bind to antibodies or to other proteins with high affinity.…”

Section: Researchmentioning

confidence: 99%

Engineering a regulatable enzyme for homogeneous immunoassays

1999

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We have engineered the phage displayed TEM-1 beta-lactamase to generate enzymes that can be used in homogeneous immunoassays because their activity can be modulated by binding to monoclonal antibodies (Mabs) raised against an unrelated protein. Random peptide libraries were genetically inserted into three loops to create hybrid enzymes with binding sites for Mabs. Insertion points were chosen to be close enough to the active site that complex formation could affect the activity. The antibiotic resistance provided by the beta-lactamase activity was used to select the clones encoding active enzymes. Biopanning of the active libraries on immobilized Mabs against the prostate specific antigen (PSA) or on streptavidin yielded enzymes with binding sites for these proteins. Their activity could be regulated by Mab or streptavidin binding. The dissociation constants of the complexes are in the 10(-9) to 10(-6) M range. In a competitive assay, PSA could be detected at a minimal concentration of 10(-9) M. The Mabs recognize mimotopes as no sequence similarity was found between inserts in regulated clones and fragments of the PSA sequence. The method can be developed to generate signaling molecules to be used for the detection of analytes in solution without identification of the epitope.

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Selection of peptides with surface affinity for α-chymotrypsin using a phage display library

Cited by 20 publications

References 20 publications

Tumor targeting with a selective gelatinase inhibitor

Tumor targeting with a selective gelatinase inhibitor

Identification of novel prostate‐specific antigen‐binding peptides modulating its enzyme activity

Engineering a regulatable enzyme for homogeneous immunoassays

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