The human body contains a million or so distinct proteins. Chemistry harbors the potential to provide ready access to these natural proteins as well as to create nonnatural ones with desirable attributes. During the previous grant period, new chemical means were discovered to manipulate protein structure and protein function.
Specific Aims. The overall goal of the proposed research is to use ideas and methods from organic chemistry and chemical biology to extend our fundamental understanding of the chemical reactivity of proteins, and to employ that understanding in meaningful ways. During the next grant period, this intent will be achieved in four Specific Aims.
Aims 1 -3 employ chemistry to effect the bioreversible modification of protein amino groups, carboxyl groups, and sulfhydryl groups.
Aim 4 integrates three state-of-the-art methods in protein chemistry (nonnatural amino acid mutagenesis, expressed protein ligation, and the traceless Staudinger ligation) to produce authentic ubiquitin conjugates and to use those conjugates to reveal key molecular aspects of protein degradation by the proteasome. Notably, the modifications in Aims 1 and 2 will provide distinct means to deliver proteins into human cells, and those in Aims 1-3 will be performed on an important tumor suppressor protein, PTEN. Significance. The results of the research proposed herein will provide new insight into the intrinsic and extrinsic chemical reactivity of proteins, as well as extend the capacity to access and manipulate proteins. The knowledge gained will have a broad and deep impact on biomedicine in this post-genomic era.

Public Health Relevance

A protein is a gene-encoded string of amino acids that folds into a three-dimensional structure. Proteins perform the molecular functions that are necessary for life, including catalysis of biochemical reactions (by enzymes), neutralization of foreign toxis (by antibodies), and stimulation of cellular activity (by hormones). The goal of this project is to develop chemical means to manipulate proteins with a precision that is unobtainable with other (e.g., genetic) methods and to endow proteins thereby with desirable attributes that could be transformative to biomedicine.

National Institute of Health (NIH)
Research Project (R01)
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Synthetic and Biological Chemistry B Study Section (SBCB)
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Fabian, Miles
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University of Wisconsin Madison
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United States
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Mix, Kalie A; Aronoff, Matthew R; Raines, Ronald T (2016) Diazo Compounds: Versatile Tools for Chemical Biology. ACS Chem Biol 11:3233-3244
Newberry, Robert W; Raines, Ronald T (2016) A prevalent intraresidue hydrogen bond stabilizes proteins. Nat Chem Biol 12:1084-1088
Gold, Brian; Aronoff, Matthew R; Raines, Ronald T (2016) Decreasing Distortion Energies without Strain: Diazo-Selective 1,3-Dipolar Cycloadditions. J Org Chem 81:5998-6006
Aronoff, Matthew R; Gold, Brian; Raines, Ronald T (2016) Rapid cycloaddition of a diazo group with an unstrained dipolarophile. Tetrahedron Lett 57:2347-2350
Gold, Brian; Aronoff, Matthew R; Raines, Ronald T (2016) 1,3-Dipolar Cycloaddition with Diazo Groups: Noncovalent Interactions Overwhelm Strain. Org Lett 18:4466-9
Aronoff, Matthew R; Gold, Brian; Raines, Ronald T (2016) 1,3-Dipolar Cycloadditions of Diazo Compounds in the Presence of Azides. Org Lett 18:1538-41
Andersen, Kristen A; Smith, Thomas P; Lomax, Jo E et al. (2016) Boronic Acid for the Traceless Delivery of Proteins into Cells. ACS Chem Biol 11:319-23
Arnold, Ulrich; Raines, Ronald T (2016) Replacing a single atom accelerates the folding of a protein and increases its thermostability. Org Biomol Chem 14:6780-5
Johnston, Sean B; Raines, Ronald T (2016) PTENpred: A Designer Protein Impact Predictor for PTEN-related Disorders. J Comput Biol 23:969-975
Johnston, Sean B; Raines, Ronald T (2015) Conformational stability and catalytic activity of PTEN variants linked to cancers and autism spectrum disorders. Biochemistry 54:1576-82

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