The human body contains two million or so distinct proteins. Chemistry harbors the potential to endow these proteins with attributes desirable for biomedicine. The proposed research takes advantage of new chemical reactivity that is applicable in a physiological context. The overall goal is to develop a facile, general means to endow native proteins with the ability to enter the cytosol of human cells. The strategy is to O-alkylate protein carboxyl groups by using a tuned diazo compound, generating esters that are analogous to those in small- molecule prodrugs. Intracellular esterases make these modifications traceless, avoiding any compromise to proper function or potential for immunogenicity. ?Protein esterification? has an uncharted landscape. Accordingly, work will begin by exploring fundamental attributes of esterified proteins, including the mechanism of cellular uptake and the enzymology of ester hydrolysis by cellular esterases in vitro and in cellulo (Aim 1). Esterification will then be used to deliver proteins for tumor suppression (Aim 2), genome editing (Aim 3), and anti-viral activity (Aim 4). The work relies on methods and ideas from organic chemistry, enzymology, chemical biology, and related fields, and is designed to establish a new paradigm for developing chemotherapeutic agents that are capable of addressing numerous unmet medical needs.

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 toxins (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 #
3R01GM044783-27S1
Application #
9892594
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Fabian, Miles
Project Start
1990-07-01
Project End
2022-06-30
Budget Start
2019-07-01
Budget End
2020-06-30
Support Year
27
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
001425594
City
Cambridge
State
MA
Country
United States
Zip Code
02142
Windsor, Ian W; Palte, Michael J; Lukesh 3rd, John C et al. (2018) Sub-picomolar Inhibition of HIV-1 Protease with a Boronic Acid. J Am Chem Soc 140:14015-14018
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
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
Mix, Kalie A; Lomax, Jo E; Raines, Ronald T (2017) Cytosolic Delivery of Proteins by Bioreversible Esterification. J Am Chem Soc 139:14396-14398
Smith, Thomas P; Windsor, Ian W; Forest, Katrina T et al. (2017) Stilbene Boronic Acids Form a Covalent Bond with Human Transthyretin and Inhibit Its Aggregation. J Med Chem 60:7820-7834
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
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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