Escherichia coli maltose-binding protein as a molecular chaperone for recombinant intracellular cytoplasmic single-chain antibodies 1 1Edited by R. Huber

Bach, Horacio; Mazor, Yariv; Shaky, Shelly; Shoham-Lev, Atar; Berdichevsky, Yevgeny; Gutnick, David L.; Benhar, Itai

doi:10.1006/jmbi.2001.4914

Cited by 164 publications

(128 citation statements)

References 74 publications

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“…HP0149 (fused either to His 6 or MBP) was toxic; therefore, the interaction that HP0149 is engaged in could not be studied (Table II). These results are in agreement with several studies describing MBP tags as an efficient tag to enhance solubility of fusion proteins (33)(34)(35). Expression and solubility results of the His 6 tag proteins are overall comparable to those reported in other expression studies (35).…”

Section: Cloning Expression and Solubility Of The Proteinssupporting

confidence: 91%

Biochemical Characterization of Protein Complexes from the Helicobacter pylori Protein Interaction Map

Terradot

Durnell

et al. 2004

Molecular & Cellular Proteomics

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We have investigated a large set of interactions from the Helicobacter pylori protein interaction map previously identified by high-throughput yeast two-hybrid (htY2H)-based methods. This study had two aims: i) to validate htY2H as a source of protein-protein interaction complexes for high-throughput biochemical and structural studies of the H. pylori interactome; and ii) to validate biochemically interactions shown by htY2H to involve components of the H. pylori type IV secretion systems. Thus, 17 interactions involving 31 proteins and protein fragments were studied, and a general strategy was designed to produce protein-interacting partners for biochemical and structural characterization. We show that htY2H is a valid source of protein-protein complexes for high-throughput proteome-scale characterization of the H. pylori interactome, because 76% of the interactions tested were confirmed biochemically. Of the interactions involving type IV secretion proteins, three could be confirmed. One interaction is between two components of the type IV secretion apparatus, ComB10 and ComB4, which are VirB10 and VirB4 homologs, respectively. Another interaction is between a type IV component (HP0525, a VirB11 homolog) and a non-type IV secretion protein (HP01451), indicating that proteins other than the core VirB (1-11)-VirD4 proteins may play a role in type IV secretion. Finally, a third interaction was biochemically confirmed between CagA, a virulence factor secreted by the type IV secretion system encoded by the Cag pathogenicity island, and a non-type IV secretion protein, HP0496.

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Section: Cloning Expression and Solubility Of The Proteinssupporting

confidence: 91%

Biochemical Characterization of Protein Complexes from the Helicobacter pylori Protein Interaction Map

Terradot

Durnell

et al. 2004

Molecular & Cellular Proteomics

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“…An unknown in these experiments is how fusion with MBP [used as a molecular chaperone to promote peptide solubility and stability (24)] might affect the presentation of the tilted peptide. MBP is a large protein with two distinct globular domains separated by a deep groove, and at its surface lie several clusters of hydrophobic residues (25).…”

Section: Discussionmentioning

confidence: 99%

Prolactin/growth hormone–derived antiangiogenic peptides highlight a potential role of tilted peptides in angiogenesis

Nguyen

Tabruyn

Lins

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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Angiogenesis is a crucial step in many pathologies, including tumor growth and metastasis. Here, we show that tilted peptides exert antiangiogenic activity. Tilted (or oblique-oriented) peptides are short peptides known to destabilize membranes and lipid cores and characterized by an asymmetric distribution of hydrophobic residues along the axis when helical. We have previously shown that 16-kDa fragments of the human prolactin͞growth hormone (PRL͞GH) family members are potent angiogenesis inhibitors. Here, we demonstrate that all these fragments possess a 14-aa sequence having the characteristics of a tilted peptide. The tilted peptides of human prolactin and human growth hormone induce endothelial cell apoptosis, inhibit endothelial cell proliferation, and inhibit capillary formation both in vitro and in vivo. These antiangiogenic effects are abolished when the peptides' hydrophobicity gradient is altered by mutation. We further demonstrate that the well known tilted peptides of simian immunodeficiency virus gp32 and Alzheimer's ␤-amyloid peptide are also angiogenesis inhibitors. Taken together, these results point to a potential new role for tilted peptides in regulating angiogenesis. 16-kDa N-terminal fragment of prolactinA ngiogenesis, the formation of new blood vessels from preexisting ones, is crucial in both health and disease. Its involvement in tumor growth and metastasis (1) makes it an important potential target for anticancer therapy. Many angiogenesis inhibitors are cryptic fragments of endogenous molecules displaying no angiogenesis-related activity [angiostatin, endostatin, platelet factor-4 (PF-4), tumstatin, and thrombospondin (2)]. From some of these inhibitors, shorter peptides retaining antiangiogenic activity have been isolated, such as the PF-4 peptide 47-70 (3), endostatin fragments 2 and 5 (amino acids 60-70 and amino acids 171-183) (4), and tumstatin peptides T3 and T7 (amino acids 69-88 and amino acids 74-98) (5). This observation suggests that the antiangiogenic regions of these peptides may be exposed in the isolated fragments but buried in the full-length proteins.The 16-kDa N-terminal fragment of prolactin (16K PRL) is antiangiogenic in vitro (6-11) and in vivo. Generation of the 16K fragment from PRL has been attributed to cathepsin D (12). Its ability to prevent angiogenesis in tumor and retinopathy mouse models has raised interest in its potential therapeutic use (13-15). We have sought to identify in human 16K PRL (16K hPRL) a peptide that might be responsible for its antiangiogenic activity. Although the 16K fragments of the other three human PRL͞GH-family members are also potently antiangiogenic (16), the sequence similarity of these fragments is low (Ϸ35% similarity between all mammalian PRL͞GH sequences). This consideration led us to seek a peculiar common structural feature rather than a similar sequence.Tilted (or oblique-oriented) peptides are short helical peptides (11 to 20 aa long) characterized by a peculiar distribution of hydrophobic residues: they are amphipathic ...

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“…40 Some of the examples given for scFvs that exhibited improved expression upon fusion to tags were already confirmed to be well-functioning intrabodies when expressed without a fusion tag. 40,48,49 Therefore, fusion to a Cκ-domain or to solubility enhancers might lead to enhanced expression or improved performance of an already confirmed functional intrabody, but this method may not be sufficient to convert all scFvs into intrabodies.…”

confidence: 99%

Targeting antibodies to the cytoplasm

Marschall

Frenzel

Schirrmann

et al. 2011

mAbs

109

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Escherichia coli maltose-binding protein as a molecular chaperone for recombinant intracellular cytoplasmic single-chain antibodies 1 1Edited by R. Huber

Cited by 164 publications

References 74 publications

Biochemical Characterization of Protein Complexes from the Helicobacter pylori Protein Interaction Map

Biochemical Characterization of Protein Complexes from the Helicobacter pylori Protein Interaction Map

Prolactin/growth hormone–derived antiangiogenic peptides highlight a potential role of tilted peptides in angiogenesis

Targeting antibodies to the cytoplasm

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