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Available in vitro and in vivo methods for verifying protein substrates for …

Biology Articles » Biochemistry » Protein Biochemistry » Farnesylation or geranylgeranylation? Efficient assays for testing protein prenylation in vitro and in vivo » Abbreviations

- Farnesylation or geranylgeranylation? Efficient assays for testing protein prenylation in vitro and in vivo

FPP, farnesylpyrophosphate; FTase, farnesyltransferase; FTI, farnesyltransferase inhibitor; GAP, GTPase activating protein; GEF, guanine nucleotide exchange factor; GFP, green fluorescent protein; GGPP, geranylgeranylpyrophosphate; GGTase1, geranylgeranyltransferase 1; GGTase2, geranylgeranyltransferase 2; GGTI, geranylgeranyltransferase inhibitor; GST, glutathione-S-transferase; HA, hemagglutinin; PBS, phosphate-buffered saline; PCR, polymerase chain reaction; PTases, prenyltransferases; PTM, posttranslational modification; SDS-PAGE, sodium dodecyl sulphate polyacrylamide gelelectrophoresis; TLC, thin layer chromatography

Authors' contributions
FE initiated this research. The experiments were performed by WB and MK. WB, MK, FP and FE conceived the work and advised modifications of experimental designs. The sequence-analytic part was carried out by SMS. WB, MK and FE contributed to the writing of the manuscript and all authors read and approved the final text.

The GFP-RhoA plasmids were kindly provided by Channing J. Der (University of North Carolina, USA). The authors received support in methodical questions of detection of lipid-modified proteins with HPLC and mass-spectrometry by Mike Ferguson, Nick Morris, Douglas Lamont and Kenny Beattie (University of Dundee). Help in establishing the biochemical laboratory was provided by Gustav Ammerer, Andreas Hartig, Franz Koller, Wolfgang Löffelhardt and Wolfgang Reiter (University Vienna) as well as Kim Nasmyth, Barry McGuinness, Thorsten Decker, Gunnar Schotta, Kerstin Wendt, Peter Steinlein and Visnja Pavicic (IMP Vienna). Josefina Pinon is thanked for editing the manuscript. The authors are grateful for financial support from Boehringer Ingelheim. Chromatography equipment was generously provided by Agilent Technologies Vienna (with support from Franz Weigang). This project has been partly funded by the Austrian Gen-AU bioinformatics integration network (BIN) sponsored by BM-BWK (grant to FE).

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