Host–Guest Complexation by β-Cyclodextrin Enhances the Solubility of an Esterified Protein
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nihms-1834418.pdf
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Accepted version
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947.98 KB
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Author(s)
Cheah, Keith M
Jun, Joomyung V
Wittrup, K Dane
Raines, Ronald T
Date Issued
August 29, 2022
Journal
Molecular Pharmaceutics
Publisher
American Chemical Society
Citation
Keith M. Cheah, Joomyung V. Jun, K. Dane Wittrup, and Ronald T. Raines
Molecular Pharmaceutics 2022 19 (11), 3869-3876.
Version
Author's final manuscript
Abstract
The carboxyl groups of a protein can be esterified by reaction with a diazo compound, 2-diazo-2-(p-methylphenyl)-N,N-dimethylacetamide. This esterification enables the entry of the protein into the cytosol of a mammalian cell, where the nascent ester groups are hydrolyzed by endogenous esterases. The low aqueous solubility of the ensuing esterified protein is, however, a major practical challenge. Solubility screening revealed that β-cyclodextrin (β-CD) is an optimal solubilizing agent for esterified green fluorescent protein (est-GFP). Its addition can increase the recovery of est-GFP by 10-fold. α-CD, γ-CD, and cucurbit-7-uril are less effective excipients. 1H NMR titration experiments revealed that β-CD encapsulates the hydrophobic tolyl group of ester conjugates with Ka = 321 M–1. Combining l-arginine and sucrose with β-CD enables the nearly quantitative recovery of est-GFP. Thus, the insolubility of esterified proteins can be overcome with excipients.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Koch Institute for Integrative Cancer Research at MIT
Massachusetts Institute of Technology. Department of Chemistry
Massachusetts Institute of Technology. Department of Biological Engineering
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DOI of Published Version
10.1021/acs.molpharmaceut.2c00368
