Detection and Quantification of Protein Post-Translational ModificationsUsing Novel Microchannel Plate Autoradiographic Imagers

摘要

It is likely that all proteins are modified post-translationally, either enzymatically or non-enzymatically. Scientists have continued to develop suitable methodologies in order to study the ever burgeoning list of protein post-translational modifications (PTMs). For the study of PTMs, one must consider suitable assays by which the target protein can be detected, the site(s) of PTM determined, the stoichiometry and stability of the PTM evaluated, and the biological outcome of the modification assessed. Historically, one of the methods of choice for identifying which protein is modified, and the site and stoichiometry of modification, has been the utilisation of commercial radiochemicals. Of these, tritium (3H) has excellent potential to facilitate analyses of biochemical reactions, but has the weakest signal strength of the commonly employed biological β-emitters. Hence development of imaging devices that are superior to conventional film autoradiography will enable a better exploitation of the universal application of 3H for measuring PTMs. Herein, we discuss the current application of film autoradiography and the advantages of using microchannel plate (MCP) autoradiographic imagers.