A mild procedure for solid phase peptide synthesis: use of fluorenylmethoxycarbonylamino-acids

Atherton, Eric; Fox, Hazel M.; Harkiss, Diana; Logan, C.; Sheppard, R. C.; Williams, Brian J.

doi:10.1039/c39780000537

Cited by 294 publications

(139 citation statements)

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“…53,54 Applied for the first time for SPPS by two different laboratories independently. 55,56 Since then, several optimized removal conditions for SPS have been described, the most relevant being: 20% piperidine in DMF, 55 which is the most common, 1-5% DBU in DMF, 57,58 morpholine-DMF (1:1) 59 or 2% HOBt, 2% hexamethyleneimine, 25% N-methylpyrrolidine in DMSO-NMP (1:1), 60 Fmoc α-amino protection has been used for the SPS of several relevant peptides using the so-called Fmoc/ t Bu strategy, the production in Tm scale of the T20 peptide being one of the most important examples. 63 Nevertheless, the low solubility of some Fmoc derivatives in the most commonly used solvents for SPPS has stimulated the search for new base-labile protecting groups.…”

Section: Protecting Groups Removed By Basementioning

confidence: 99%

Amino Acid-Protecting Groups

2009

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Section: Protecting Groups Removed By Basementioning

confidence: 99%

Amino Acid-Protecting Groups

2009

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“…These so-called large immunophilins (FKBP52, FKBP51, and cyclophilin 40) have been found in complexes with the chaperone Hsp90 and certain client proteins such as the steroid hormone receptors in their inactive states (11)(12)(13). For steroid hormone receptor activation, several Hsp90 complexes of different compositions seem to be needed (14).…”

Section: Conversion Of Prolyl Peptide Bonds From Trans-to Cis-prolinesmentioning

confidence: 99%

Localization of the Chaperone Domain of FKBP52

Pirkl¹,

Fischer²,

Modrow³

et al. 2001

Journal of Biological Chemistry

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FKBP52, a multidomain peptidyl prolyl cis/transisomerase (PPIase), is found in complex with the chaperone Hsp90 and the co-chaperone p23. It displays both PPIase and chaperone activity in vitro. To localize these two activities to specific regions of the protein, we created and analyzed a set of fragments of FKBP52. The PPIase activity toward both peptides and proteins is confined entirely to domain 1 (amino acids 1-148). The chaperone activity, however, resides in the C-terminal part of FKBP52, mainly in the region between amino acids 264 and 400 (domain 3). Interestingly, this domain also contains the tetratricopeptide repeats, which are responsible for the binding to C-terminal amino acids of Hsp90. Competition assays with a C-terminal Hsp90 peptide suggest that the non-native protein and Hsp90 are bound by different regions within this domain.

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“…A linear peptide was synthesized using conventional bmoc chemistry [19,201 and cleaved from the resin with the cysteine residues blocked with Acm groups. This procedure allowed more control over the formation of disulphide bonds and the purification of the peptide to homogeneity before Acm cleavage.…”

Section: Resultsmentioning

confidence: 99%

Synthesis and characterization of μ‐conotoxin IIIa

Becker

Atherton

Gordon

1989

European Journal of Biochemistry

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p-Conotoxin 111 a, a voltage-dependent sodium channel neurotoxin, has been synthesised using solid-phase peptide synthesis employing 9-fluorenylmethoxycarbonyl chemistry. After cleavage from the resin, the peptide was isolated by reverse-phase HPLC and then the six acetamidomethyl groups were removed by treatment with mercuric acetate. The reduced product so formed was purified by reverse-phase HPLC. Protocols were developed to optimize the oxidation of the cysteine residues to form disulphide bonds. Protocols employed using air oxidation together with 2-mercaptoethanol were the most effective. As complete oxidation was never obtained the oxidised peptide was purified by reverse-phase HPLC. The acitivity of our products was monitored using [3H]saxitoxin binding to eel membranes. The oxidised product was able to completely block [3H]saxitoxin binding in a competitive manner. Lineweaver-Burke analysis of ['H]saxitoxin binding gave a K, of 1.5 nM, 1Cs0 was determined as 26.6 nM. It was also shown that the pure synthetic p-conotoxin IIIa had the same retention time on reversephase HPLC as the natural conotoxin IIIa. Thus an active toxin has been synthesised that can be used to probe sodium channels.Neurotoxins from various natural sources have been essential tools for labelling and physiologically manipulating the sodium channel [l]. Current understanding of toxin binding to the sodium channel implies the existence of five or more discrete toxin-binding sites. These binding sites are (a) the tetrodotoxin/saxitoxin-binding site, at which ligands inhibit ion transport, (b) the alkaloid-binding site at which toxins like batrachotoxin and veratridine bind to produce persistent activation of the sodium channel, (c) the North African scorpion a-toxinlsea anemone toxin-binding site where these reagents act to remove inactivation, (d) the American scorpion j-toxin site at which toxins mediate a shift in inactivation and (e) the ciguatoxin/ptychodiscus sitc; reagents binding here cause repetitive firing and persistent activation [2, 31.More recently a series of sodium-channel-specific toxins derived from the marine gastropod family Conidne have been described [4]. These purified polypeptide toxins from Conus geographus, the so-called p-conotoxins (also called geographutoxins) have a common structure and elicit a blockade of sodium currents by high-affinity binding to the tetrodotoxin/saxitoxin-binding site [5,6]. These toxins can block electric eel Electrophorus electricus and skeletal muscle sodium channels but not channels from brain, heart or nerve tissues [5, 7. 81. p-Conotoxins can be used to electrophysiologically distinguish between the different sodium channels that are observed during muscle development in a manner to the more widely described tetrodotoxin sensitivity [9, lo]. The amino acid sequences have been described and reveal that these toxins contain the rare amino acids hydroxyproline together with many cysteine residues [5, 61. The latter observation suggests that these toxins have a multi-looped structu...

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A mild procedure for solid phase peptide synthesis: use of fluorenylmethoxycarbonylamino-acids

Abstract: Use of base-labile N-fluorenylmethoxycarbonylamino-acids, t-butyl based side chain protecting groups, and a p-alkoxybenzyl ester resin linkage provides a simple, rapid, and exceptionally mild strategy in solid phase peptide synthesis.

Cited by 294 publications

References 0 publications

Amino Acid-Protecting Groups

Amino Acid-Protecting Groups

Localization of the Chaperone Domain of FKBP52

Synthesis and characterization of μ‐conotoxin IIIa

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