This Core provides peptide, protein, antibody and RNA reagents that support HIV/Host structural and mechanistic studies within the CHEETAH Center. Bacterial expression systems will be the workhorse for producing recombinant proteins without complex folding/post-translational modification requirements, while eukaryotic expression systems will be employed to produce more complex proteins. Uniformly glycosylated or deglycosylated proteins will be made using advanced tissue culture and enzymatic approaches.1-3 Membrane proteins will be produced by coupling eukaryotic expression with comprehensive detergent screening protocols.4 Peptide synthesis will be employed for the production of ligands, inhibitors, and protein fragments. Larger synthetic peptides/proteins will be produced using native chemical ligation to enable the introduction of site-specific labels and modifications.5 Finally, an automated high-throughput phage display screening and characterization platform will be used to produce Fab antibody fragments for use as crystallography chaperones, cryo-EM fiducials, biochemistry/cell biology tools, and to stabilize/trap heterogeneous protein complexes in specific conformations for structural studies.6
| Wagner, Jonathan M; Christensen, Devin E; Bhattacharya, Akash et al. (2018) General Model for Retroviral Capsid Pattern Recognition by TRIM5 Proteins. J Virol 92: |
| Donaldson, G P; Ladinsky, M S; Yu, K B et al. (2018) Gut microbiota utilize immunoglobulin A for mucosal colonization. Science 360:795-800 |
| Bailey, Lucas J; Sheehy, Kimberly M; Dominik, Pawel K et al. (2018) Locking the Elbow: Improved Antibody Fab Fragments as Chaperones for Structure Determination. J Mol Biol 430:337-347 |
| Pak, Alexander J; Voth, Gregory A (2018) Advances in coarse-grained modeling of macromolecular complexes. Curr Opin Struct Biol 52:119-126 |
| Redman, Joseph S; Francis, J Nicholas; Marquardt, Robert et al. (2018) Pharmacokinetic and Chemical Synthesis Optimization of a Potent d-Peptide HIV Entry Inhibitor Suitable for Extended-Release Delivery. Mol Pharm 15:1169-1179 |
| Larsen, Kevin P; Mathiharan, Yamuna Kalyani; Kappel, Kalli et al. (2018) Architecture of an HIV-1 reverse transcriptase initiation complex. Nature 557:118-122 |
| Carter, Stephen D; Mageswaran, Shrawan K; Farino, Zachary J et al. (2018) Distinguishing signal from autofluorescence in cryogenic correlated light and electron microscopy of mammalian cells. J Struct Biol 201:15-25 |
| Shepherd, Jason D (2018) Arc - An endogenous neuronal retrovirus? Semin Cell Dev Biol 77:73-78 |
| Coey, Aaron T; Larsen, Kevin P; Choi, Junhong et al. (2018) Dynamic Interplay of RNA and Protein in the Human Immunodeficiency Virus-1 Reverse Transcription Initiation Complex. J Mol Biol 430:5137-5150 |
| Swulius, Matthew T; Nguyen, Lam T; Ladinsky, Mark S et al. (2018) Structure of the fission yeast actomyosin ring during constriction. Proc Natl Acad Sci U S A 115:E1455-E1464 |
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