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.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM044783-22A1
Application #
8766779
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Fabian, Miles
Project Start
1990-07-01
Project End
2018-04-30
Budget Start
2014-08-01
Budget End
2015-04-30
Support Year
22
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Biochemistry
Type
Earth Sciences/Resources
DUNS #
City
Madison
State
WI
Country
United States
Zip Code
53715
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Chyan, Wen; Kilgore, Henry R; Raines, Ronald T (2018) Cytosolic Uptake of Large Monofunctionalized Dextrans. Bioconjug Chem 29:1942-1949
Chyan, Wen; Raines, Ronald T (2018) Enzyme-Activated Fluorogenic Probes for Live-Cell and in Vivo Imaging. ACS Chem Biol 13:1810-1823
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Hoang, Trish T; Smith, Thomas P; Raines, Ronald T (2017) A Boronic Acid Conjugate of Angiogenin that Shows ROS-Responsive Neuroprotective Activity. Angew Chem Int Ed Engl 56:2619-2622
Burke, Eileen G; Gold, Brian; Hoang, Trish T et al. (2017) Fine-Tuning Strain and Electronic Activation of Strain-Promoted 1,3-Dipolar Cycloadditions with Endocyclic Sulfamates in SNO-OCTs. J Am Chem Soc 139:8029-8037
Chyan, Wen; Kilgore, Henry R; Gold, Brian et al. (2017) Electronic and Steric Optimization of Fluorogenic Probes for Biomolecular Imaging. J Org Chem 82:4297-4304
Newberry, Robert W; Raines, Ronald T (2016) A prevalent intraresidue hydrogen bond stabilizes proteins. Nat Chem Biol 12:1084-1088
Mix, Kalie A; Aronoff, Matthew R; Raines, Ronald T (2016) Diazo Compounds: Versatile Tools for Chemical Biology. ACS Chem Biol 11:3233-3244

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