Natasha T Snider1, M Bishr Omary2. 1. Department of Cell Biology and Physiology, University of North Carolina, Chapel Hill, North Carolina, USA. Electronic address: ntsnider@med.unc.edu. 2. Department of Molecular & Integrative Physiology, Department of Medicine, University of Michigan, Ann Arbor, Michigan, USA; VA Ann Arbor Healthcare System, Ann Arbor, Michigan, USA.
Abstract
Intermediate filament (IF) proteins are known to be regulated by a number of posttranslational modifications (PTMs). Phosphorylation is the best-studied IF PTM, whereas ubiquitination, sumoylation, acetylation, glycosylation, ADP-ribosylation, farnesylation, and transamidation are less understood in functional terms but are known to regulate specific IFs under various contexts. The number and diversity of IF PTMs is certain to grow along with rapid advances in proteomic technologies. Therefore, the need for a greater understanding of the implications of PTMs to the structure, organization, and function of the IF cytoskeleton has become more apparent with the increased availability of data from global profiling studies of normal and diseased specimens. This chapter will provide information on established methods for the isolation and monitoring of IF PTMs along with the key reagents that are necessary to carry out these experiments.
Intermediate filament (IF) proteins are known to be regulated by a number of posttranslational modifications (PTMs). Phosphorylation is the best-studied IF PTM, whereas ubiquitination, sumoylation, acetylation, n class="Chemical">glycosylationpan>, ADP-ribosylationpan>, farnpan>esylationpan>, anpan>d tranpan>samidationpan> are less unpan>derstood inpan> funpan>ctionpan>al terms but are knpan>own to regulate specific IFs unpan>der various conpan>texts. The number anpan>d diversity of IF PTMs is certainpan> to grow alonpan>g with rapid advanpan>ces inpan> proteomic technpan>ologies. Therefore, the need for a greater unpan>derstanpan>dinpan>g of the implicationpan>s of PTMs to the structure, organpan>izationpan>, anpan>d funpan>ctionpan> of the IF cytoskeletonpan> has become more apparenpan>t with the inpan>creased availability of data from global profilinpan>g studies of normal anpan>d diseased specimenpan>s. This chapter will provide inpan>formationpan> onpan> established methods for the isolationpan> anpan>d monpan>itorinpan>g of IF PTMs alonpan>g with the key reagenpan>ts that are necessary to carry out these experimenpan>ts.
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