Preparation and application of the 5-(4-(9-fluorenylmethyloxycarbonyl)aminomethyl-3,5-dimethoxyphenoxy)-valeric acid (PAL) handle for the solid-phase synthesis of C-terminal peptide amides under mild conditions

Alberício, Fernando; Kneib-Cordonier, Nancy; Biancalana, Sara; Gera, Lajos; Masada, R. Irene; Hudson, Derek; Bárány, George

doi:10.1021/jo00299a011

Cited by 319 publications

(187 citation statements)

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“…The p53 peptides [sequences 319-347, 319-351, 319-360, 332-360, 360-393, Ac-Cys-Gly-Gly-319-351 (AC), and Bz-319-351-Gly-GlyCys (BC)] were synthesized by the solid-phase method, using fluoren-9-ylmethoxycarbonyl (Fmoc) chemistry on an Applied Biosystems 430A peptide synthesizer. Cleavage of the peptides from the resin and removal of the side-chain protecting groups were accomplished by using reagent K (13 …”

Section: Methodsmentioning

confidence: 99%

Specific sequences from the carboxyl terminus of human p53 gene product form anti-parallel tetramers in solution.

Sakamoto

Lewis

Kodama

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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Human p53 is a tumor-suppressor gene product associated with control of the cell cycle and with growth suppression, and it is known to form homotetramers in solution. To investigate the relationship of structure to tetramerization, nine peptides corresponding to carboxyl-terminal sequences in human p53 were chemically synthesized, and their equilibrium associative properties were determined by analytical ultracentrifugation. Secondary structure, as determined by circular dichroism measurements, was correlated with oligomerization properties of each peptide. The sedimentation profiles of peptides 319-393 and 319-360 fit a two-state model of peptide monomers in equilibrium with peptide tetramers. Successive deletion of amino-and carboxyl-terminal residues from 319-360 reduced tetramer formation. Further, substitution of alanine for Leu-323, Tyr-327, and Leu-330 abolished tetramerization. Circular dichroism studies showed that peptide 319-351 had the highest a-helix content, while the other peptides that did not form tetramers had low helical structure. These studies define a minimal region and identify certain critical residues involved in tetramerization. Cross-linking studies between monomer units in the tetramer suggest that the helices adopt an anti-parallel arrangement. We propose that conformational shifts in the helical structure of the p53 tetramerization domain result in a repositioning of subunits relative to one another. This repositioning provides an explanation relating conformational changes at the carboxyl terminus with changes in sequence-specific DNA binding by the highly conserved central domain.The p53 gene product is a phosphoprotein which has been implicated in the regulation of cell proliferation. Functional inactivation of the protein either by association with viral proteins (1) or, more frequently, by point mutations, as found in many human tumors, leads to cellular transformation (2). Wild-type p53 exhibits trans-activation of transcriptional activity which is thought to mediate its anti-proliferation activity (3). At the biological level, p53 undergoes oligomerization, and it can adopt two distinct conformational states, with low or high affinity for site-specific DNA binding (4). Many of the p53 mutants associated with human tumors appear to be incapable of binding DNA and can be distinguished from wild-type p53 by their oligomerization state and by their "mutant" immunoreactivity (5-7).Recent structure-function analyses of p53 have mapped the trans-activation domain to the 42 amino (N)-terminal amino acids (8). This region is highly susceptible to proteolytic digestion and may be loosely folded in the absence of ligand (9). Deletion analysis has shown that residues 90-286 form a stable domain which binds DNA with specificity (4).Subtilisin treatment of the p53 protein generates a 191-amino acid protease-resistant monomeric fragment from the midsection that corresponds to a highly conserved region in which the majority of missense mutations occur (9, 10).We have a limited unders...

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Section: Methodsmentioning

confidence: 99%

Specific sequences from the carboxyl terminus of human p53 gene product form anti-parallel tetramers in solution.

Sakamoto

Lewis

Kodama

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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“…Petroleum ether refers to the fraction with a boiling point of 40-60 1C. 3,5-Diethoxyphenol, 26 2,6-dimethoxy-4-hydroxybenzaldehyde (3), 27,28 and (E)-1-chloro-4-(2-tetrahydropyranyloxy)-2-butene (11) 29 were prepared according to known procedures. TentaGel S-Br (90 mm, 0.26 mmol di Br per gram) was purchased from Fluka (Buchs, Switzerland).…”

Section: Experimental Procedures Materialsmentioning

confidence: 99%

“…The benzylamine derivatives 1 and 2 were prepared according to Scheme 3, starting from the known benzaldehyde 3 27,28 or the new benzaldehyde 4 obtained through the Vilsmeier formylation of 3,5-diethoxyphenol 26 (Supplementary Information).…”

Section: Synthesis Of Benzylamine Derivatives 1 Andmentioning

confidence: 99%

Synthesis and evaluation of resins bearing substrate-like inhibitor functions for capturing copper amine oxidases

Pocci

Alfei

Castellaro

et al. 2013

Polym J

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The aim of this work was to make new tools available for investigating the interactions between enzymes and substrate-like inhibitors in copper amine oxidases (CAOs), a class of enzymes that controls important cellular processes, such as the crosslinking of elastin and collagen, cell proliferation and the regulation of intracellular polyamines. Starting from previous work, we synthesized the poly(N,N-dimethylacrylamide)-based resin R2 and the new TentaGel resins T3 and T4 obtained by ether bonding CAO substrate-like inhibitor moieties onto commercial TentaGel S-Br, which contains bromomethyl groups susceptible to nucleophilic substitution reactions. We used polyacrylamide gel electrophoresis (PAGE) experiments to determine the capability of the prepared resins to capture plasma amine oxidase (PAO) and diamine oxidase (DAO), members of the CAO family. The poly(N,N-dimethylacrylamide)-based resin R2 was able to block PAO and DAO, being the first insoluble polymeric material capable of capturing enzymes of the CAO family. Keywords: copper amine oxidases; 2,6-dialkoxybenzylamines; electrophoresis; functionalization; tentagel resins INTRODUCTIONCopper amine oxidases (CAOs, EC 1.4.3.6) are ubiquitous enzymes found in prokaryotes and eukaryotes, humans included, where they control important cellular processes, such as the crosslinking of elastin and collagen, cell proliferation and the regulation of intracellular polyamines. With the exception of lysyl oxidase (32 kDa), CAOs are homodimers with subunit molecular weights between 70 kDa and 95 kDa, and with one copper ion and a quinone cofactor per subunit. CAOs are also characterized by the presence of cysteine residues and a variable percentage of carbohydrates, depending on the enzyme source.The enzymatic reaction results in the deamination of aminomethyl substrates to produce aldehydes, ammonia and reduced molecular oxygen in the form of hydrogen peroxide.Knowledge concerning the physiological role, mechanism of action 1-4 and numerous X-ray structures 5-14 of these enzymes, as well as methods for protein expression of CAOs, 15 making larger quantities of protein samples available, which is useful for various experiments, has grown progressively in the past two decades.Remarkably, one of the X-ray structures indicates that the complex between 2-hydrazinopyridine, a non-substrate-like, nonselective, carbonyl group scavenger inhibitor, and Escherichia coli amine oxidase results in the formation of an imine double bond between the NH 2 of the inhibitor and the C-5 carbonyl group of the cofactor 2,4,5-trihydroxyphenylalanine quinone. 16

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“…[86] Thus, numerous linkers which allow for post-modification of the resin and for "mild" cleavage conditions are the current ideal. Established acid-labile compounds in SPPS are, for example, the WANG linker, [88] the SASRIN (super acid sensitive resin) linker, [89][90] the PAL (peptide amide linker), [91][92] and the RINK type linkers [93] ( Figure 14, a-d). Besides acid-labile linkers, many other linkers exist which can, for instance, be released by electrophilic, nucleophilic, oxidative, reductive, photo-induced, or metal-assisted cleavage.…”

Section: Ii23 Cleavable Linkersmentioning

confidence: 99%

Purification of Peptides in High-Complexity Arrays

Schirwitz

2013

Springer Theses

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Compared to the human genome, which is formed by approximately 25,000 genes, the human proteome comprises over a million functional proteins and thus represents a much higher diversity. Although genetic research provides valuable information on the proteins which can be translated, a great task in the future will be the exploration of the entire protein interaction network. Understanding protein interactions will greatly help scientists to identify the mechanisms behind fatal diseases such as cancer, AIDS, and tuberculosis and, hopefully, provide new

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Preparation and application of the 5-(4-(9-fluorenylmethyloxycarbonyl)aminomethyl-3,5-dimethoxyphenoxy)-valeric acid (PAL) handle for the solid-phase synthesis of C-terminal peptide amides under mild conditions

Cited by 319 publications

References 0 publications

Specific sequences from the carboxyl terminus of human p53 gene product form anti-parallel tetramers in solution.

Specific sequences from the carboxyl terminus of human p53 gene product form anti-parallel tetramers in solution.

Synthesis and evaluation of resins bearing substrate-like inhibitor functions for capturing copper amine oxidases

Purification of Peptides in High-Complexity Arrays

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