Chemical Synthesis of Membrane Proteins

Schmid, Alanca; Becker, Christian F. W.

doi:10.1002/9783527823567.ch16

Cited by 5 publications

(7 citation statements)

References 121 publications

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“…Removable backbone or side‐chain modifications based on oligoethylene glycol [43] or on poly‐Lys/Arg tags. [ 44 , 45 , 46 , 47 , 48 , 49 ] have been previously used to increase the solubility of peptides. Photocleavable linker 14 was synthesized in 7 steps with an overall yield of 42 % on a multigram scale (Scheme 3 ; for details on the synthesis see Supporting Information Scheme S1).…”

Section: Resultsmentioning

confidence: 99%

Semisynthesis of Homogeneous, Active Granulocyte Colony‐Stimulating Factor Glycoforms

et al. 2022

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Granulocyte colony stimulating factor (G-CSF) is a cytokine used to treat neutropenia. Different glycosylated and non-glycosylated variants of G-CSF for therapeutic application are currently generated by recombinant expression. Here, we describe our approaches to establish a first semisynthesis strategy to access the aglycone and O-glycoforms of G-CSF, thereby enabling the preparation of selectively and homogeneously post-translationally modified variants of this important cytokine. Eventually, we succeeded by combining selenocysteine ligation of a recombinantly produced N-terminal segment with a synthetic C-terminal part, transiently equipped with a side-chain-linked, photocleavable PEG moiety, at low concentration. The transient PEGylation enabled quantitative enzymatic elongation of the carbohydrate at Thr133. Overall, we were able to significantly reduce the problems related to the low solubility and the tendency to aggregate of the two protein segments, which allowed the preparation of four G-CSF variants that were successfully folded and demonstrated biological activity in cell proliferation assays.

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Section: Resultsmentioning

confidence: 99%

Semisynthesis of Homogeneous, Active Granulocyte Colony‐Stimulating Factor Glycoforms

et al. 2022

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“…HPLC), thus making it more difficult to remove by‐products during purification steps. For this reason, access to full‐length lipidated proteins usually necessitates the assembly of multiple peptide and lipopeptide fragments using peptide ligation chemistry, or the use of transient solubilizing modifications that are also employed in the synthesis of membrane peptides and proteins [117] . In this context, the use of alternative ligation strategies such as the STL, DSL, and direct aminolysis methods have become common, as palmitoyl thioesters are often unstable in the presence of thiol additives used to enhance the rates of NCL reactions [118–120] …”

Section: Figurementioning

confidence: 99%

Chemical Synthesis and Semisynthesis of Lipidated Proteins

et al. 2022

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Lipidation is a ubiquitous modification of peptides and proteins that can occur either co‐ or post‐translationally. An array of different lipid classes can adorn proteins and has been shown to influence a number of crucial biological activities, including the regulation of signaling, cell–cell adhesion events, and the anchoring of proteins to lipid rafts and phospholipid membranes. Whereas nature employs a range of enzymes to install lipid modifications onto proteins, the use of these for the chemoenzymatic generation of lipidated proteins is often inefficient or impractical. An alternative is to harness the power of modern synthetic and semisynthetic technologies to access lipid‐modified proteins in a pure and homogeneously modified form. This Review aims to highlight significant advances in the development of lipidation and ligation chemistry and their implementation in the synthesis and semisynthesis of homogeneous lipidated proteins that have enabled the influence of these modifications on protein structure and function to be uncovered.

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“…Ausd iesem Grund ist es normalerweise notwendig, mehrere Peptid-und Lipopeptidfragmente mittels Ligationschemie zu einem Protein zusammenzufügen oder transiente Solubisierungs-Tags zu nutzen, wie sie auch bei der Synthese von Membranpeptiden und -proteinen eingesetzt werden. [117] In diesem Kontext werden häufig alternative Ligationsstrategien, wie beispielsweise STL, DSL und direkte Aminolyse,g enutzt, da Palmitoylthioester in Gegenwart von Thioladditiven, die genutzt werden, um die Geschwindigkeit von NCL-Reaktionen zu erhçhen, instabil sind. [118][119][120] Bis heute wurden zahlreiche palmitoylierte Proteine durch chemische Totalsynthese erhalten.…”

Section: Synthese Und Semisynthese Von Palmitoyliertenunclassified

Chemische Synthese und Semisynthese von lipidierten Proteinen

Hanna¹,

Kriegesmann

Dowman

et al. 2022

Angewandte Chemie

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Lipidierung ist eine ubiquitäre Modifikation von Peptiden und Proteinen, die entweder co‐ oder posttranslational auftreten kann. Für die Vielzahl von Lipidklassen wurde gezeigt, dass diese viele entscheidende biologische Aktivitäten, z. B. die Regulierung der Signalweiterleitung, Zell‐Zell‐Adhäsion sowie die Anlagerung von Proteinen an Lipid‐Rafts und Phospholipidmembranen, beeinflussen. Während die Natur Enzyme nutzt, um Lipidmodifikationen in Proteine einzubringen, ist ihre Nutzung für die chemoenzymatische Herstellung von lipidierten Proteinen häufig ineffizient. Eine Alternative ist die Kombination moderner synthetischer und semisynthetischer Techniken, um lipidierte Proteine in reiner und homogen modifizierter Form zu erhalten. Dieser Aufsatz erörtert Fortschritte in der Entwicklung der Lipidierungs‐ und Ligationschemie und deren Anwendung in der Synthese und Semisynthese homogen lipidierter Proteine, die es ermöglichen, den Einfluss dieser Modifikationen auf die Proteinstruktur und ‐funktion zu untersuchen.

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Chemical Synthesis of Membrane Proteins

Cited by 5 publications

References 121 publications

Semisynthesis of Homogeneous, Active Granulocyte Colony‐Stimulating Factor Glycoforms

Semisynthesis of Homogeneous, Active Granulocyte Colony‐Stimulating Factor Glycoforms

Chemical Synthesis and Semisynthesis of Lipidated Proteins

Chemische Synthese und Semisynthese von lipidierten Proteinen

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