A Lewis Acid-Catalyzed Procedure for the Conversion of 2-Methoxyethoxymethyl Ethers to Carboxylic Esters

A procedure for the conversion of 2-methoxy-ethoxymethyl (MEM) ethers to carboxylic esters employed ferric chloride (0.4 equivalents) and a carboxylic anhydride (14 equivalents) and exhibited selectivity for the MEM ether functionality in the presence of benzyl ethers.     

Fast Photochemical Oxidation of Proteins (FPOP) Maps the Epitope of EGFR Binding to Adnectin

Epitope mapping is an important tool for the development of monoclonal antibodies, mAbs, as therapeutic drugs. Recently, a class of therapeutic mAb alternatives, adnectins, has been developed as targeted biologics. They are derived from the 10th type III domain of human fibronectin (¹⁰Fn3). A common approach to map the epitope binding of these therapeutic proteins to their binding partners is X-ray crystallography. Although the crystal structure is known for Adnectin 1 binding to human epidermal growth factor receptor (EGFR), we seek to determine complementary binding in solution and to test the efficacy of footprinting for this purpose. As a relatively new tool in structural biology and complementary to X-ray crystallography, protein footprinting coupled with mass spectrometry is promising for protein–protein interaction studies. We report here the use of fast photochemical oxidation of proteins (FPOP) coupled with MS to map the epitope of EGFR-Adnectin 1 at both the peptide and amino-acid residue levels. The data correlate well with the previously determined epitopes from the crystal structure and are consistent with HDX MS data, which are presented in an accompanying paper. The FPOP-determined binding interface involves various amino-acid and peptide regions near the N terminus of EGFR. The outcome adds credibility to oxidative labeling by FPOP for epitope mapping and motivates more applications in the therapeutic protein area as a stand-alone method or in conjunction with X-ray crystallography, NMR, site-directed mutagenesis, and other orthogonal methods. Figure

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What is a protein crystal? Can we apply the terminology of classical industrial crystallization to them?

The nature of protein crystals is discussed. Since protein crystals contain not only proteins but also other substances the usage of conventional terms (of industrial crystallization) of naming such crystals is questioned. The proof that there are other components than the protein itself in a protein crystal is given. It is suggested to use a procedure like in biotechnology to guarantee the production of the right protein crystals according to the PAT concept. It is suggested not to use other established names like “hydrates”, “solvates”, “but crystals for proteins since the definitions do not fit the nature of the protein crystals.