L. Deterding, K. Tomer
2008
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Abstract
In an effort to gain tertiary structural information of proteins, chemical modification of surface exposed residues and chemical crosslinking have been used in combination with mass spectrometry. For this work, the acetylation of lysine residues was used as a chemical modification reagent. The lysine residues that are determined to be acetylated are, thus, assumed to be surface accessible. The chemical crosslinker DTSSP, 3, 3′-dithiobis-[sulfosuccinimidylpropionate], was used for the crosslinking experiments. DTSSP is a homobifunctional amine reactive crosslinker that is cleavable. Therefore, an aliquot of the crosslinked protein was then subjected to conditions that would result in cleavage of the linker. After performing all modification and crosslinking experiments, the protein was digested and analyzed by both MALDI and LC/ESI mass spectrometry. For proof-of-principle, bovine β-lactoglobulin was used for these analyses, and the results are compared to the X-ray crystal structure of the protein. As an extension of this work, these biochemical and mass spectrometric techniques have been applied to a protein of unknown structure.