Isolation of novel single-chain Cro proteins targeted for binding to the bcl-2 transcription initiation site by repertoire selection and subunit combinatorics
2005 (English)In: Protein Engineering Design & Selection, ISSN 1741-0126, E-ISSN 1741-0134, Vol. 18, no 11, 537-546 p.Article in journal (Refereed) Published
New designed DNA-binding proteins may be recruited to act as transcriptional regulators and could provide new therapeutic agents in the treatment of genetic disorders such as cancer. We have isolated tailored DNA-binding proteins selected for affinity to a region spanning the transcription initiation site of the human bcl-2 gene. The proteins were derived from a single-chain derivative of the lambda Cro protein (scCro), randomly mutated in its recognition helices to construct libraries of protein variants of distinct DNA-binding properties. By phage display-afforded affinity selections combined with recombination of shuffled subunits, protein variants were isolated, which displayed high affinity for the target bcl-2 sequence, as determined by electrophoretic mobility shift and biosensor assays. The proteins analyzed were moderately sequence-specific but provide a starting point for further maturation of desired function.
Place, publisher, year, edition, pages
2005. Vol. 18, no 11, 537-546 p.
phage display, protein–DNA interactions, repressor, single-chain Cro, subunit shuffling
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:uu:diva-74639DOI: 10.1093/protein/gzi058PubMedID: 16186141OAI: oai:DiVA.org:uu-74639DiVA: diva2:102549