Journal article
In vitro and cellular assays for palmitoyl acyltransferases using fluorescent lipidated peptides
Methods (San Diego, Calif.), Vol.40(2), pp.166-170
2006
Handle:
https://hdl.handle.net/2376/115696
PMCID: PMC2892119
PMID: 17012028
Abstract
Protein palmitoylation is emerging as an important post-translational modification in development as well as in the establishment and progression of diseases such as cancer. This chapter describes the use of fluorescent lipidated peptides to characterize palmitoyl acyltransferase (PAT) activities
in vitro and in intact cells. The peptides mimic two motifs that are enzymatically palmitoylated, i.e. C-terminal farnesyl and N-terminal myristoyl sequences. These substrate peptides can be separated from the palmitoylated product peptides by reversed-phase HPLC, detected and quantified by the fluorescence of their NBD label. Through these methods, the activities of PATs toward these alternate substrates in isolated membranes or intact cells can be quantified. The
in vitro assay has been used to characterize human PATs and to identify inhibitors of these enzymes. The cellular assay has been useful in elucidating the kinetics of protein palmitoylation by PATs
in situ, and the sub-cellular distribution of the palmitoylated products.
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Details
- Title
- In vitro and cellular assays for palmitoyl acyltransferases using fluorescent lipidated peptides
- Creators
- Charles E Ducker - Apogee Biotechnology Corporation, P.O. Box 916, Hershey, PA 17033, USAJeremiah M Draper - Department of Pharmacology, Penn State College of Medicine, Hershey, PA 17033, USAZuping Xia - Department of Pharmaceutical Sciences, Medical University of South Carolina, Charleston, SC 29425, USACharles D Smith - Apogee Biotechnology Corporation, P.O. Box 916, Hershey, PA 17033, USA
- Publication Details
- Methods (San Diego, Calif.), Vol.40(2), pp.166-170
- Academic Unit
- Pharmacy and Pharmaceutical Sciences, College of
- Publisher
- Elsevier Inc
- Identifiers
- 99900547512901842
- Language
- English
- Resource Type
- Journal article