Assays of protein palmitoylation.

@article{citeulike:2332867, abstract = {Protein palmitoylation plays an important role in the structure and function of a wide array of proteins. Unlike other lipid modifications, protein palmitoylation is highly dynamic and cycles of palmitoylation and depalmitoylation can regulate protein function and localization. The dynamic nature of palmitoylation is poorly resolved because of limitations in assay methods. Here, we discuss various methods that can be used to measure protein palmitoylation and identify sites of palmitoylation. We describe new methodology based on "fatty acyl exchange labeling" in which palmitate is removed via hydroxylamine-mediated cleavage of the palmitoyl-thioester bond and then exchanged with a sulfhydryl-specific labeling compound. The techniques are highly sensitive and allow for quantitative estimates of palmitoylation. Unlike other techniques used to assay posttranslational modifications, the techniques we have developed can label all sites of modification with a variety of probes, radiolabeled or non-radioactive, and can be used to assay the palmitoylation of proteins from tissue samples.}, address = {Departments of Neurobiology, Pharmacology and Physiology, University of Chicago, Chicago, IL 60637, USA.}, author = {Drisdel, R. C. and Alexander, J. K. and Sayeed, A. and Green, W. N. }, citeulike-article-id = {2332867}, doi = {10.1016/j.ymeth.2006.04.015}, issn = {1046-2023}, journal = {Methods}, month = {October}, number = {2}, pages = {127--134}, posted-at = {2008-02-05 02:05:49}, priority = {2}, title = {Assays of protein palmitoylation.}, url = {http://dx.doi.org/10.1016/j.ymeth.2006.04.015}, volume = {40}, year = {2006} }

TY - JOUR ID - citeulike:2332867 L3 - citeulike-article-id:2332867 TI - Assays of protein palmitoylation. JF - Methods VL - 40 IS - 2 SP - 127 EP - 134 AD - Departments of Neurobiology, Pharmacology and Physiology, University of Chicago, Chicago, IL 60637, USA. SN - 1046-2023 N2 - Protein palmitoylation plays an important role in the structure and function of a wide array of proteins. Unlike other lipid modifications, protein palmitoylation is highly dynamic and cycles of palmitoylation and depalmitoylation can regulate protein function and localization. The dynamic nature of palmitoylation is poorly resolved because of limitations in assay methods. Here, we discuss various methods that can be used to measure protein palmitoylation and identify sites of palmitoylation. We describe new methodology based on "fatty acyl exchange labeling" in which palmitate is removed via hydroxylamine-mediated cleavage of the palmitoyl-thioester bond and then exchanged with a sulfhydryl-specific labeling compound. The techniques are highly sensitive and allow for quantitative estimates of palmitoylation. Unlike other techniques used to assay posttranslational modifications, the techniques we have developed can label all sites of modification with a variety of probes, radiolabeled or non-radioactive, and can be used to assay the palmitoylation of proteins from tissue samples. AU - Drisdel, RC AU - Alexander, JK AU - Sayeed, A AU - Green, WN PY - 2006/10// UR - http://dx.doi.org/10.1016/j.ymeth.2006.04.015 ER -