Studies of Surface Preparation for the Fluorosequencing of Peptides.

Silica passivating agents have shown great success in minimizing nonspecific protein binding to glass surfaces for imaging and microscopy applications. Amine-derivatized surfaces are commonly used in conjugation with amide coupling agents to immobilize peptides/proteins through C-terminal or side-chain carboxylic acids. In the case of the single-molecule fluorosequencing of peptides, attachment occurs via the C-terminus and nonspecific surface binding has previously been a source of error in peptide identification.

Mechanistic studies of a "Declick" reaction.

A kinetic analysis of a "declick" reaction is described. Compound , previously reported to couple an amine and a thiol ( "click") under mild aqueous conditions to create , undergoes release of the unaltered coupling partners upon triggering with dithiothreitol (). In the study reported herein various aniline derivatives possessing para-electron donating and withdrawing groups were used as the amines.