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. Hydrus 2d 3d Crack Tattoo · VPN Unlimited 6.0 .A versatile one-pot approach to target aldehyde-substituted bioorthogonal lysine in proteome analysis.
A variety of bioconjugation methods are commonly used for the analyses of protein structures, interactions, and functions. However, these techniques are mainly restricted to the reversible covalent modification of functional groups, such as hydroxyl or amine groups. Therefore, various proteins with post-translational modifications, such as aldehyde groups, cannot be efficiently analyzed. In this study, we developed a one-pot lysine-aldehyde modification strategy for the versatile and specific site-specific labeling of target aldehyde-containing proteins. To reduce the chemoselectivity of the aldehyde-lysine conjugation, an amine-selective N-hydroxysuccinimide-mediated labeling protocol and a copper-free, nitrogen-selective carbonyl-selective azide-alkyne cycloaddition (Cu-free N(3)AAC) were newly developed to enable efficient, specific and efficient labeling. To overcome the unspecific modification of protein structures, the lysine-aldehyde was conjugated with the guanidinium groups of arginine residues to detect and remove excess reactive lysine-aldehyde. This novel modification method was successfully applied to the specific detection of α-synuclein, which was overexpressed in pathologic conditions. The method was also useful for the analysis of post-translational modifications of α-synuclein. Furthermore, the method can be applied to the in vivo analyses of α-synuclein. Considering that the currently available method of labeling lysine-aldehydes requires complicated synthetic steps, our newly developed technique can be applied to a variety of cell and tissue analyses.Q:
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