Probing biomolecular interactions of glutathione transferase M2-2 by using peptide phage display.
The interactions between biomolecules and human glutathione transferase M2-2 (GST M2-2) were probed by using 9- and 15-mer combinatorial peptide libraries displayed on phage. The peptide libraries were based on random DNA sequences fused to gIII, a gene that expresses a phage coat protein and thus causes the peptides to be displayed on the surface of phage particles. A peptide sequence was enriched through binding to GST M2-2, which indicated a successful selection. Binding studies with the peptide displayed on phage showed binding specificity. The sequence of the peptide had similarities to segments of proteins in the Swiss-Prot Database, to c-Jun N-terminal kinase (JNK), and to the protein Bcl3. JNK is linked to the regulation of the transcription factor AP-1. Use of cell-based assays of the transcriptional activity of AP-1 allowed a novel coactivation function of GST M2-2 to be demonstrated. Specificity in the activation was indicated by the lack of effect of GST A1-1. No coactivator function of GST M2-2 could be demonstrated in assays with Bcl3. These results suggest that GST M2-2 has biological roles in addition to catalysis of detoxication reactions, and demonstrate the potential of phage display in functional genomics research.
Edalat
Maryam
M
Department of Biochemistry, Biomedical Center Box 576, Uppsala University, 751 23 Uppsala, Sweden.
Pettersson
Sven
S
Persson
Mats A A
MA
Mannervik
Bengt
B
D000081242:B-Cell Lymphoma 3 Protein; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D015336:Molecular Probe Techniques; D009842:Oligopeptides / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000187:drug effects / Q000235:genetics; D014157:Transcription Factors; D015533:Transcriptional Activation / Q000187:drug effects; D014407:Tumor Cells, Cultured
D000081242:B-Cell Lymphoma 3 Protein; C000631344:BCL3 protein, human; D007527:Isoenzymes; D009842:Oligopeptides; D019151:Peptide Library; D011518:Proto-Oncogene Proteins; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D005982:Glutathione Transferase; C412602:glutathione S-transferase Mu 2; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases