Professor, Biological Sciences
My lab studies the folding and design of proteins, enzymes, vaccine antigens, and biosensors. When I came here in 1999, I was a bioinformaticist and my lab worked exclusively in computational biology. Since then, we have evolved into a mostly experimental lab, with a little computation still mixed in. Our early work included algorithm development in protein structural bioinformatics, including structure prediction, contact map prediction, local structure prediction, hidden Markov models, sequence alignment, molecular surface area calculation, torsion space molecular dynamics, simplified representations for molecular dynamics, bioinformatics-driven force fields, structure-based alignment, and models for folding pathways. We proposed the "phone cord effect" to explain superhelicity in alpha helical crossovers.
Starting about ten years ago, we began doing experimental work on green fluorescent protein (GFP) with the help of molecular biologist Prof. Donna E. Crone. We created a non-circularly permuted "re-wired" GFP, a permuted and truncated "leave-one-out" GFP, a variant that folds faster and more efficiently, and a GFP-base biosensor. We inserted strategically-placed disulfides to test specific hypotheses for the folding pathway of GFP and reasoned out the presence of two specific folding intermediates. More recently we are studying the formation of the fluorescent chromophore and the dependence of GFP fluorescence on specific residues and on thermodynamic stability.
The principle thrust of the lab is to develop the leave-one-out (LOO) method for GFP-based biosensors. By combining computational design and high throughput screening, we find sets of mutations that complement a specific bound peptide and allow the truncated LOO-GFP to fold and glow only in the presence of a target protein which has been unfolded to expose that sequence. A biosensor for detection of H5N1 influenza virus hemagglutinin was published in 2015. We are collaborating on the development of a sensor for dengue virus and putting those sensors on protein fibers.
We are also engaged in a collaborative effort to develop a contraceptive vaccine. The vaccine will produce temporary and reversible infertility. Instead of having to take an action to be protected, you will be protected by default, and need to take an action to be temporarilly fertile again. The vaccine targets the sperm-specific calcium channel CatSper, which is required for sperm hyperactive motility and therefore for fertility. Sperm antigens are placed on the surface of non-infectious virus-like particles (VLP) for vaccination.
This work was made possible by our collaborators, the Center for Computational Innovation (CCI), and Center for Biotechnology and Interdisciplinary Sciences (CBIS), and by grants from the NSF, NIH, Rosetta Commons, and a private foundation.
BA. Carleton College, Northfield MN.
Ph.D. University of California, San Diego
- protein folding
- protein design
- Kizer, M., Huntress, I. D., Walcott, B., Fraser, K., Bystroff, C., & Wang, X. (2019). The complex between a multi-crossover DNA nanostructure, PX-DNA, and T7 endonuclease I. Biochemistry. Mar 12;58(10):1332-1342.
- Bystroff, C. (2018). Intramembranal disulfide cross-linking elucidates the super-quaternary structure of mammalian CatSpers. Reproductive Biology, 18(1), 76-82.
- Hooper WF, Walcott BD, Wang X, Bystroff C. (2018) Fast design of arbitrary length loops in proteins using InteractiveRosetta. BMC Bioinformatics, 19(1), 337.
- Bystroff C (2018) Will an anti-sperm contraceptive vaccine gain public acceptance? Obstetrics and Gynecology Reports. Volume 2(3): 1-1. ISSN: 2515-2955
- Banerjee S, Schenkelberg CD, Jordan TB, Reimertz JM, Crone EE, Crone DE, & Bystroff C. (2017). Mispacking and the Fitness Landscape of the Green Fluorescent Protein Chromophore Milieu. Biochemistry, 56(5), 736-747.
- Shirke AN, Basore D, Holton S, Su A, Baugh E, Butterfoss GL, Makhatadze G, Bystroff C, Gross RA. (2016) Influence of surface charge, binding site residues and glycosylation on Thielavia terrestris cutinase biochemical characteristics. Applied microbiology and biotechnology. 2016 Jan 13:1-2
- Schenkelberg CD, Bystroff C. (2016) Protein backbone ensemble generation explores the local structural space of unseen natural homologs. Bioinformatics. 2016 Jan 18:btw001.
- Huang Y-M, Banerjee S, Crone DE, Buck PM, Schenkelberg CD, Pitman DJ & Bystroff C. (2015) Computationally designed biosensors based on leave-one-out green fluorescent protein. Biochemistry 54(40):6263-73.