Robert E. Campbell "Engineering fluorescent proteins into new biotools: manipulation of protein surfaces, topology, and chromophores"
Robert E. Campbell
Assistant Professor
University of Alberta
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At the heart of the Campbell group's research program lies the extraordinarily useful family of fluorescent proteins (FPs) which includes the green FP (GFP) from Aequorea jellyfish and its numerous homologues of various color from coral. All FPs share the ability to autonomously generate a visible wavelength fluorophore from a sequence of 3 amino acids located in the center of their barrel-like structures. FPs are credited with sparking the explosive and continuing growth in the popularity of live cell fluorescence microscopy; culminating in the 2008 Nobel Prize in Chemistry being awarded to 3 pioneers in this area; Shimomura, Chalfie, and Tsien.
Since its inception in 2003, the Campbell research group has worked to mould (that is, engineer) FPs into improved tools for addressing fundamental questions in life science. For example, we have developed several improved alternatives to some of the traditionally preferred hues of FPs [1,2,3] and exploited their improved properties in new applications. Some of our novel applications include the use of FPs for investigating peptide structure in vivo [4], dual-FRET imaging of caspase activation during apoptosis [5], and a FP-based assay of nucleoside transporter activity (with Joe Casey) [6]. In this seminar I will discuss some of our most recent efforts to create FP variants that have physical or spectral properties that are either improved relative to existing variants or are completely novel and have not been previously reported. As the title of this seminar indicates, this discussion will focus on 3 aspects of FPs; chain topology, surface properties, and chromophore chemistry. Specifically, I will present our most recent progress in the following areas: creation of circularly permuted red FPs through manipulation of topology; engineering of monomeric and heterodimeric FPs through manipulation of surface properties; and engineering of photostable and photconvertable FPs by manipulation of chromophore chemistry and environment.