The membranes of a cell are hotspots of biological activity. In turn, many diseases proceed through action near or within cellular membranes. Members of the peripheral membrane class of proteins are cytosolic until targeted to the membrane, either through covalent or non-covalent means. We are interested in understanding these proteins at atomic resolution. This information helps us gain a deeper understanding of these proteins and is used to inform rational inhibitor design efforts. The ultimate goal of the Fuglestad Lab is to inhibit peripheral membrane proteins for chemical biology investigations and to develop drug candidates to treat diseases such as cancer and cardiovascular disorders.
Understanding the interactions between peripheral membrane proteins and lipids is of paramount importance in strategizing therapeutics. We use a number of models of biological membranes to probe the interactions between proteins and cellular membranes. Of particular interest in the Fuglestad Lab is leveraging reverse micelles as membrane mimics, which has several advantages over other more commonly used membrane models. We employ a multitude of techniques to study protein/membrane interactions including protein nuclear magnetic resonance (NMR) spectroscopy, molecular dynamics simulation, and small-angle scattering, among others.
Fragment screening and inhibitor design
Rational design of inhibitors uses information about the structure and function of the protein target of interest. Fragment-based approaches have recently come to prominence in inhibitor design. This involves screening proteins for small inhibitor building blocks rather than larger drug-like molecules. Despite its tremendous promise, one instance where fragment screening generally fails is in the detection of inhibitor building blocks for ‘smooth’ proteins lacking natural ligand-binding pockets. We employ a recently developed technology that takes advantage of nanoscale confinement of proteins within reverse micelles to allow detection of fragment binding to smooth proteins. The information gained from these screens is used in highly collaborative efforts in inhibitor development for the ultimate goal of obtaining chemical biology tools and drug leads.
Labrecque CL, Fuglestad B. Electrostatic Drivers of GPx4 Interactions with Membrane, Lipids, and DNA. Biochemistry, Epub ahead of print, Sept. 7, 2021. doi.org/10.1021/acs.biochem.1c00492
O'Brien ES, Fuglestad B, Lessen HJ, Stetz MA, Lin DW, Marques BS, Gupta K, Fleming KG, Wand AJ. Membrane Proteins Have Distinct Fast Internal Motion and Residual Conformational Entropy. Angew Chem Int Ed Engl. 2020, 59, 11108-11114
Fuglestad B, Kerstetter NE, and Wand, AJ. Site-resolved and quantitative characterization of very weak protein-ligand interactions. ACS Chem Biol. 2019 Jul 19;14(7):1398-1402.
Fuglestad B, Gupta K, Wand AJ, Sharp KA. Water loading driven size, shape, and composition of cetyltrimethylammonium/hexanol/pentane reverse micelles. J Colloid Interface Sci. 2019 Mar 22;540:207-217.
Fuglestad B, Marques BS, Jorge C, Kerstetter NE, Valentine KG, Wand AJ. Reverse Micelle Encapsulation of Proteins for NMR Spectroscopy. Methods Enzymol. 2019;615:43-75.
O'Brien ES, Lin DW, Fuglestad B, Stetz MA, Gosse T, Tommos C, Wand AJ. Improving yields of deuterated, methyl labeled protein by growing in H(2)O. J Biomol NMR. 2018 Aug;71(4):263-273.
Fuglestad B, Stetz MA, Belnavis Z, Wand AJ. Solution NMR investigation of the response of the lactose repressor core domain dimer to hydrostatic pressure Biophys Chem. 2017 Dec;231:39-44.
Handley LD*, Fuglestad B*, Stearns K, Tonelli M, Fenwick RB, Markwick PR, Komives EA. NMR reveals a dynamic allosteric pathway in thrombin. Sci Rep. 2017 Jan 6;7:39575.
Fuglestad B, Gupta K, Wand AJ, Sharp KA. Characterization of Cetyltrimethylammonium Bromide/Hexanol Reverse Micelles by Experimentally Benchmarked Molecular Dynamics Simulations. Langmuir. 2016 Feb 23;32(7):1674-84.
O'Brien ES, Nucci NV, Fuglestad B, Tommos C, Wand AJ. Defining the Apoptotic Trigger: the interaction of cytochrome c and cardiolipin. J Biol Chem. 2015 Dec 25;290(52):30879-87.
Nucci NV, Fuglestad B, Athanasoula EA, Wand AJ. Role of cavities and hydration in the pressure unfolding of T4 lysozyme. Proc Natl Acad Sci U S A. 2014 Sep 23;111(38):13846-51.
Fuglestad B*, Gasper PM*, McCammon JA, Markwick PR, Komives EA. Correlated motions and residual frustration in thrombin. J Phys Chem B. 2013 Oct 24;117(42):12857-63.
Gasper PM, Fuglestad B, Komives EA, Markwick PR, McCammon JA. Allosteric networks in thrombin distinguish procoagulant vs. anticoagulant activities. Proc Natl Acad Sci U S A. 2012 Dec 26;109(52):21216-22.
Fuglestad B, Gasper PM, Tonelli M, McCammon JA, Markwick PR, Komives EA. The dynamic structure of thrombin in solution. Biophys J. 2012 Jul 3;103(1):79-88.