title: Versatile Interacting Peptide (VIP) Tags for Labeling Proteins with Bright Chemical Reporters.
abstract: Fluorescence microscopy is an essential tool for the biosciences, enabling the direct observation of proteins in their cellular environment. New methods that facilitate attachment of photostable synthetic fluorophores with genetic specificity are needed to advance the frontiers of biological imaging. Here, we describe a new set of small, selective, genetically encoded tags for proteins based on a heterodimeric coiled-coil interaction between two peptides: CoilY and CoilZ. Proteins expressed as a fusion to CoilZ were selectively labeled with the complementary CoilY fluorescent probe peptide. Fluorophore-labeled target proteins were readily detected in cell lysates with high specificity and sensitivity. We found that these versatile interacting peptide (VIP) tags allowed rapid and specific delivery of bright organic dyes or quantum dots to proteins displayed on living cells. Additionally, we validated that either CoilY or CoilZ could serve as the VIP tag, which enabled us to observe two distinct cell-surface protein targets with this one heterodimeric pair.
authors.affiliation: Department of Biomedical Engineering, Oregon Health & Science University, 2730 Moody Avenue (CL3B), Portland, OR, 97201, USA.
authors.forename: Hannah K
authors.initials: HK
authors.lastname: Zane
authors.affiliation: Department of Biomedical Engineering, Oregon Health & Science University, 2730 Moody Avenue (CL3B), Portland, OR, 97201, USA.
authors.forename: Julia K
authors.identifier: 0000-0001-9375-6947
authors.initials: JK
authors.lastname: Doh
authors.affiliation: Department of Cell, Developmental and Cancer Biology, Oregon Health & Science University, 2730 Moody Avenue (CL3B), Portland, OR, 97201, USA.
authors.forename: Caroline A
authors.initials: CA
authors.lastname: Enns
authors.affiliation: Department of Biomedical Engineering, Oregon Health & Science University, 2730 Moody Avenue (CL3B), Portland, OR, 97201, USA.
authors.forename: Kimberly E
authors.identifier: 0000-0002-2303-8779
authors.initials: KE
authors.lastname: Beatty
mesh_terms: D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D018613:Microscopy, Confocal; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides; D049452:Green Fluorescent Proteins
keywords: coiled coils; conjugation; fluorescent probes; microscopy; protein engineering