Increasing evidence suggests that ribosomes actively regulate protein synthesis. However, much of this evidence is indirect, leaving this layer of gene regulation largely unexplored, in part due to methodological limitations. Indeed, we review evidence demonstrating that commonly used methods, such as transcriptomics, are inadequate because the variability in mRNAs coding for ribosomal proteins (RP) does not necessarily correspond to RP variability. Thus protein remodeling of ribosomes should be investigated by methods that allow direct quantification of RPs, ideally of isolated ribosomes. We review such methods, with emphasis on their biases and approaches to control these biases. We argue that using multiple complementary methods can help reduce the danger of interpreting reproducible systematic biases as evidence for ribosome remodeling.
slavovLab@slavov_n Enabling quantitative DIA analysis of samples labeled with isobaric tags will be a big step in further expanding the power of DIA analysis.
It certainly holds much promise for single-cell analysis methods using the isobaric carrier approach t.co/R80UFNC2uk