Biomedical subjects

M Brickner

Publications and source records attributed to M Brickner.

4 recordsLinked to original sources

PubMed1998

Uncoiling c-Jun coiled coils: inhibitory effects of truncated Fos peptides on Jun dimerization and DNA binding in vitro.

c-Jun is an oncoprotein that comprises a portion of the AP-1 transcription factor and belongs to the basic-leucine zipper (bZIP) DNA binding protein family. Using peptides derived from the leucine zipper region of Fos, we have developed agents that inhibit Jun's DNA binding in the low micromolar range. Fos peptides that were effective inhibitors in the DNA binding assay were also found to inhibit cellular Jun binding to an AP-1 site in a luciferase reporter plasmid in MCF-7 cells. Size exclusion studies confirmed that peptides that inhibit the DNA binding of Jun also inhibit its dimerization. These peptides were found to have a cytotoxic effect on the MCF-7 cell line when delivered with the transfecting agent Tfx-50, possibly due to their role as transcription factor regulators.

Amino Acid Sequence↗

PubMed1998

Inhibiting the dimeric restriction endonuclease EcoRI using interfacial helical peptides.

BACKGROUND: Many enzymes are active only in a dimeric form, including a variety of type II restriction endonucleases. Disruption of subunit interactions is therefore a potential method for multimeric enzyme inhibition. EcoRI is a homodimeric restriction endonuclease, the dimeric interface of which consists of a four-helix bundle. We set out to design helical peptides to interact with this interface and block dimer formation, thus rendering EcoRI inactive. RESULTS: Here we describe two synthetic, helical peptides based on the interfacial region of EcoRI. Both peptides inhibit the enzyme, but the peptide derived from the alpha 4 helix of EcoRI had both a higher helical content and better efficacy than a variant peptide, alpha 4(Leu), that has three Ile-->Leu mutations (IC50 values of 27 microM and 90 microM, and helical contents of 29% and 10%, respectively). Size-exclusion chromatography confirmed that the alpha 4 peptide disrupted dimerization of EcoRI, and circular dichroism indicated that EcoRI remained folded upon binding to alpha 4. Inhibition with alpha 4 and alpha 4(Leu) was shown to be specific for EcoRI, as the dimeric restriction enzyme PvuII was not affected by the peptides. CONCLUSIONS: Interfacial peptide inhibitors of the dimeric EcoRI were obtained that both inhibit dimerization and endonuclease activity. The peptide sequence with a preference for a helical conformation was a more effective inhibitor, presumably because the more preorganized state enhanced interactions with the helical interface of EcoRI. The specific nature of this endonuclease-peptide interaction was also confirmed. The potential of this strategy for inhibiting other enzyme classes is currently being addressed.

Chromatography, Gel↗

PubMed1998

Inhibiting the assembly of protein-protein interfaces.

Protein-protein association is found throughout mechanisms of cellular growth and differentiation, and viral replication. Inhibiting the assembly of protein complexes, therefore, presents itself as a novel means of inhibition for a wide variety of cellular and viral events. Peptides and small molecules that modify the overall quaternary structure of a selection of receptor-ligand interactions and oligomeric viral enzymes have been developed recently.

HIV↗

PubMed1982

Different difficulty manipulations interact differently with task emphasis: evidence for multiple resources.

To test the notion of multiple resources, a two-dimensional pursuit tracking task was paired with a letter-typing task, the difficulty of which was manipulated by varying cognitive (size of stimulus set) and motor (repetitiveness of finger chords) factors. In addition, task priority was manipulated. The latter factor had a large effect on the performance of the two tasks, which indicates that they compete for resources. Both types of typing difficulty manipulations affected typing performance, but only motor difficulty interacted with priorities. Since difficulty manipulations that tap resources common to both tasks are predicted to interact with priorities, the results are interpreted to indicate that in joint performance, typing and tracking compete mainly for motor-related concept, the letter-typing task is argued to require at least two kinds of resources.

Adult↗