Prenylation methods

Overmeyer, J., Erdman, R., and Maltese, W. (1998). Membrane targeting via protein prenylation. Methods Mol. Biol. 88, 249-263. 

The ferf-prenylation method has subsequently been applied by several groups to the synthesis of other natural products [101,102], and was also successfully applied to an indole without a 3-substituent [103]. 

As an alternative strategy, prelipidated building blocks can be used to generate prenylated peptides (Scheme 8). Thereby advantage is taken of the stability of the thioether linkage that offers the possibility of applying, for example, the Fmoc strategy for such peptides. Short N-Ras C-termini, for example, are readily available via this method. ... 

Two major synthetic strategies have been applied for the synthesis of S-prenylated peptides as shown in Scheme 1. In the first approach (method A), S-alkylation is performed on the final peptide in a protected or unprotected form.13641 In method B, which has produced better results in terms of overall yield, thiol alkylation is carried out on the cysteine residue or on a small peptide fragment and then the S-prenylated synthons are coupled to the remaining peptide fragment to produce the final products.[45 501 Most of the procedures reported for the synthesis of farnesylated peptide derivatives, can also be applied to the synthesis of geranylgeranylated peptides. 

An alternative and possibly more simple approach to S-prenylation is based on S-alkylation of suitably protected or unprotected peptides. This method is widely applied particularly when small amounts of S-prenylated peptides are required, whereby preparative RP-HPLC is generally applied for the isolation of the lipopeptides. The S-alkylation methods used are generally those described above. Whilst with suitably protected peptides solubility represents a major problem, which can be bypassed by S-alkylation of the peptide on resin, 61 with unprotected peptides a very efficient tuning of the reaction conditions is required... 

Recent developments in delayed extraction TOF allow higher resolving power and mass accuracy, and this method in the reflector mode has been used for accurate measurement of the mass of several compounds including two prenylated flavonoids. However, the performance of the MALDI-TOF instrument was not better than those of the FAB and FT-ICR MS instruments, and insufficient to give acceptable accuracy for literature reporting. 

Fukai, T. and Nomura, T., New NMR structure determination methods for prenylated phenols, Basic Life Sciences, 66 (Plant Polyphenols 2), 259, 1999. 

We have demonstrated that this insect cell-free protein synthesis system is one of the most effective protein synthesis systems among those based on animal extracts (2). Furthermore, it has the potential to perform eukaryote-specific protein modifications such as protein W-myristoylation and prenylation (3, 4). Thus, we expect that the insect cell-free protein synthesis system will be a useful method for target protein production in the reverse chemical genetics era, as well as for postgenomic studies. In this chapter, we describe standard protocols to synthesize proteins of interest using the insect cell-free protein synthesis system. 

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