Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences

Coin, Irene; Beyermann, Michael; Bienert, Michael

doi:10.1038/nprot.2007.454

Cited by 489 publications

(404 citation statements)

References 59 publications

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“…Depsi-peptide Aβ 1-42 was synthesized as previously described (47,48). At variance with the native peptide, the depsi-peptide is highly soluble and it has a much lower propensity to aggregate, thus preventing the spontaneous formation of seeds in the solution (49,50).…”

Section: Methodsmentioning

confidence: 99%

Synthetic amyloid-β oligomers impair long-term memory independently of cellular prion protein

Balducci¹,

Beeg

Stravalaci

et al. 2010

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

437

441

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Inability to form new memories is an early clinical sign of Alzheimer's disease (AD). There is ample evidence that the amyloid-β (Aβ) peptide plays a key role in the pathogenesis of this disorder. Soluble, bio-derived oligomers of Aβ are proposed as the key mediators of synaptic and cognitive dysfunction, but more tractable models of Aβ−mediated cognitive impairment are needed. Here we report that, in mice, acute intracerebroventricular injections of synthetic Aβ 1-42 oligomers impaired consolidation of the long-term recognition memory, whereas mature Aβ 1-42 fibrils and freshly dissolved peptide did not. The deficit induced by oligomers was reversible and was prevented by an anti-Aβ antibody. It has been suggested that the cellular prion protein (PrP C ) mediates the impairment of synaptic plasticity induced by Aβ. We confirmed that Aβ 1-42 oligomers interact with PrP C , with nanomolar affinity. However, PrP-expressing and PrP knock-out mice were equally susceptible to this impairment. These data suggest that Aβ 1-42 oligomers are responsible for cognitive impairment in AD and that PrP C is not required.Alzheimer | neurotoxicity | object recognition test | surface plasmon resonance | protein aggregation

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Section: Methodsmentioning

confidence: 99%

Synthetic amyloid-β oligomers impair long-term memory independently of cellular prion protein

Balducci¹,

Beeg

Stravalaci

et al. 2010

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

437

441

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“…Since that time, peptide synthesis has undergone several developments, beginning as solution phase reactions and eventually evolving in 1963 with Merrifield's introduction of solid resins in peptide synthesis (2), more commonly referred to as solid phase peptide synthesis (SPPS). Although this technique has undergone a dynamic maturation over the past few decades, it is still regarded as expensive and inelegant (3,4). Furthermore, there are still several challenges that are not likely to be overcome without a significant change to the synthesis approach, beyond inherent challenges such as, for example, solubility of reagents.…”

mentioning

confidence: 99%

Efficient and directed peptide bond formation in the gas phase via ion/ion reactions

McGee

McLuckey

2014

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

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Amide linkages are among the most important chemical bonds in living systems, constituting the connections between amino acids in peptides and proteins. We demonstrate the controlled formation of amide bonds between amino acids or peptides in the gas phase using ion/ion reactions in a mass spectrometer. Individual amino acids or peptides can be prepared as reagents by (i) incorporating gas phaselabile protecting groups to silence otherwise reactive functional groups, such as the N terminus; (ii) converting the carboxyl groups to the active ester of N-hydroxysuccinimide; and (iii) incorporating a charge site. Protonation renders basic sites (nucleophiles) unreactive toward the N-hydroxysuccinimide ester reagents, resulting in sites with the greatest gas phase basicities being, in large part, unreactive. The N-terminal amines of most naturally occurring amino acids have lower gas phase basicities than the side chains of the basic amino acids (i.e., those of histidine, lysine, or arginine). Therefore, reagents may be directed to the N terminus of an existing "anchor" peptide to form an amide bond by protonating the anchor peptide's basic residues, while leaving the N-terminal amine unprotonated and therefore reactive. Reaction efficiencies of greater than 30% have been observed. We propose this method as a step toward the controlled synthesis of peptides in the gas phase.peptide ligation | ion chemistry T he formation of amide bonds between amino acids holds widespread interest as it is relevant to the origin of life. Although the process by which these early peptides were formed remains an open question, laboratory-based approaches for peptide synthesis have been in place for over a century. The first controlled synthesis of diglycine via hydrolysis of diketopiperazine by Fisher in 1901 (1) marked the beginning of a long path of development of peptide synthesis approaches. Since that time, peptide synthesis has undergone several developments, beginning as solution phase reactions and eventually evolving in 1963 with Merrifield's introduction of solid resins in peptide synthesis (2), more commonly referred to as solid phase peptide synthesis (SPPS). Although this technique has undergone a dynamic maturation over the past few decades, it is still regarded as expensive and inelegant (3, 4). Furthermore, there are still several challenges that are not likely to be overcome without a significant change to the synthesis approach, beyond inherent challenges such as, for example, solubility of reagents. SPPS requires significant use of protecting and deprotecting agents, cleaving agents, and other solvents for washing, drying, and dissolving the newly formed peptides. This method quickly becomes a rather wasteful approach when considering the sacrifices made to produce just a single amide bond (3). Alternative synthesis approaches, such as chemical ligation (5, 6) and molecular machine-mediated peptide synthesis (7), offer more elegant approaches despite maintaining several of the previously mentioned requirements.Amide b...

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“…In contrast, CPDY is characterized as an exopeptidase that typically cleaves only amide bonds at C-termini. Therefore, CPDY-catalyzed polymerization avoids the hydrolysis of the polypeptide backbones of pre-existing polypeptides, resulting in efficient suppression of the broadening of the molecular weight distribution of the products that Methyl, [49,54] ethyl, [50][51][52][53][54][55][56] benzyl [54] Proteinase I, [49] papain (13)(14)(15)(16)(17)(18) [50][51][52][53][54][55][56] Glycine Methyl, [54] ethyl, [49,51,54,55] benzyl [54] Proteinase I, [49] papain (10)(11)(12)(13)(14)(15)(16)(17)(18)(19) [51,54,55] l-Leucine…”

Section: Materials and General Protocolsmentioning

confidence: 99%

Chemoenzymatic Synthesis of Polypeptides for Use as Functional and Structural Materials

Tsuchiya

Numata

2017

Macromolecular Bioscience

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Polypeptides inspired by the natural functional and structural proteins present in living systems are promising materials for various fields in terms of their versatile functionality and physical properties. Designing and synthesizing mimetic sequences of specific peptide motifs in proteins are important for exploring the functionality of natural proteins. Chemoenzymatic polymerization, which utilizes aminolysis (i.e., the reverse reaction of hydrolysis catalyzed by proteases), is a useful technique for synthesizing artificial polypeptide materials and has several advantages, including facile synthesis protocols, environmental friendliness, scalability, and atom economy. In this review, recent progress in chemoenzymatic polypeptide synthesis for the production of functional and structural materials for various applications is summarized in conjunction with the current status of technical challenges in the field.

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Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences

Cited by 489 publications

References 59 publications

Synthetic amyloid-β oligomers impair long-term memory independently of cellular prion protein

Synthetic amyloid-β oligomers impair long-term memory independently of cellular prion protein

Efficient and directed peptide bond formation in the gas phase via ion/ion reactions

Chemoenzymatic Synthesis of Polypeptides for Use as Functional and Structural Materials

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