title: Expedient chemical synthesis of 75mer DNA binding domain of MafA: an insight on its binding to insulin enhancer.
abstract: An expedient chemical synthesis of a 75mer peptide corresponding to the DNA binding domain (DBD, 227-301) of the human MafA leucine zipper transcription factor is reported. The application of microwave-assisted solid phase peptide synthesis (MW-SPPS) with a protocol modified respect to the standard one allowed obtaining the desired 75mer peptide in a short time with high quantity and optimal purity. MW-SPPS methodology was thus demonstrated as a valuable alternative to recombinant methods to obtain protein domains. Considering that recent findings suggest an involvement of MafA in the pathogenesis of diabetes mellitus, we also performed circular dichroism studies both on DBD folding and its interaction with MafA recognition element (MARE) on insulin enhancer. From our results, it was evicted that a disorder to order transition occurs after DBD interaction with insulin MARE which is mediated by specific structural elements on the N-terminus of the DBD.
authors.affiliation: Dipartimento di Scienze Molecolari Applicate ai Biosistemi, Sezione Chimica Organica, A. Marchesini, Università degli Studi di Milano, via Venezian 21, 20133 Milan, Italy. sara.pellegrino@unimi.it
authors.forename: Sara
authors.initials: S
authors.lastname: Pellegrino
authors.forename: Chiara
authors.initials: C
authors.lastname: Annoni
authors.forename: Alessandro
authors.initials: A
authors.lastname: Contini
authors.forename: Francesca
authors.initials: F
authors.lastname: Clerici
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Gelmi
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004742:Enhancer Elements, Genetic; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D016350:Leucine Zippers / Q000235:genetics*; D051473:Maf Transcription Factors, Large / Q000138:chemical synthesis* / Q000378:metabolism*; D008872:Microwaves; D008969:Molecular Sequence Data; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D060327:Solid-Phase Synthesis Techniques
chemical_list: D007328:Insulin; C485159:MAFA protein, human; D051473:Maf Transcription Factors, Large