Synthesis of cleavable peptides with authentic C-termini: an application for fully automated SPOT synthesis

Characterization of a Putative Phosphorylation Switch: Adaptation of SPOT Synthesis to Analyze PDZ Domain Regulation Mechanisms

“…Solutions of Fmoc-amino acid-OH (0.4 m) activated with 1,1'-carbonyldiimidazole (CDI, 3 equiv) [33] in DMF were spotted on the membrane (4 coupling, 15 min reaction time each). Proline, tyrosine, and glutamine were activated with 1,1'-carbonyldi(1,2,4-triazole) (CDT).…”

“…Proline, tyrosine, and glutamine were activated with 1,1'-carbonyldi(1,2,4-triazole) (CDT). [33] The Fmoc group was removed from the spots, and the sequences of the peptides were completed by the standard SPOT synthesis protocol, [22] followed by a N-terminal tag with b-alanine. For standard SPOT synthesis Fmoc-aa-Opfp were used with the following side chain protection: E-, D-(OtBu); C,-S-, T-, Y-(tBu); K-, W-(Boc); N-, Q-, H-(Trt); R-(Pbf).…”

Mapping Putative Contact Sites Between Subunits in a Bacterial ATP-binding Cassette (ABC) Transporter by Synthetic Peptide Libraries

“…Nonetheless, it was demonstrated that the coupling efficiency of aliphatic and hydrophobic amino acids is satisfactory. 18,25 Furthermore and as demonstrated recently by the successful SPOT synthesis of several 32meric leucine zipper analogs, amphipathic helical peptides can be synthesized with good yields , and MalFMalG(F403Q), respectively, were grown in LB medium (10 ml) supplemented with ampicillin (0.1 mg/ml) and chloramphenicol (0.02 mg/ml) at 37°C to late exponential phase. Subsequently, cells were desintegrated by passage through a French press and fractions corresponding to low speed pellet (LSP), cytosol (Cy) and cytoplasmic membrane vesicles (CM) were obtained as described.…”

Cellulose Functionalysed with Grafted Oligopeptides

“…N-methyl-imidazole (NMI) [19]. There are also recommendations suggesting the use of 1,1'-carbonyl-di-(1,2,4-triazole) (CDT) instead of CDI in order to reduce the risk of the deprotection of Fmoc-amino acids during the coupling reaction [20]. Since the early reports by Frank, cellulose has found widespread application as a support in the synthesis of peptides and oligonucleotides.…”