Substitutional landscape of a split fluorescent protein fragment using high-density peptide microarrays.

ABSTRACT: Split fluorescent proteins have wide applicability as biosensors for protein-protein interactions, genetically encoded tags for protein detection and localization, as well as fusion partners in super-resolution microscopy. We have here established and validated a novel platform for functional analysis of leave-one-out split fluorescent proteins (LOO-FPs) in high throughput and with rapid turnover. We have screened more than 12,000 variants of the beta-strand split fragment using high-density peptide microarrays for binding and functional complementation in Green Fluorescent Protein. We studied the effect of peptide length and the effect of different linkers to the solid support. We further mapped the effect of all possible amino acid substitutions on each position as well as in the context

SUBMITTER: Antonescu ON 

PROVIDER: S-EPMC7857580 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature