Photocontrol of Protein Activity Mediated by the Cleavage Reaction of a Split Intein

Binschik, Jens; Zettler, J. L.; Mootz, Henning D.

doi:10.1002/ange.201007078

Cited by 5 publications

(4 citation statements)

References 27 publications

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“…Photocontrollable formation of the model maltose binding protein by protein splicing was measured by a change in the fluorescence anisotropy arising from the labeling of the C intein with a fluorophore. Mootz and co‐workers synthesized an intein with an NV ‐caged diamino propionic acid, the isostere of cysteine 131. By applying prothrombin as the extein in this system, they could induce coagulation in human blood plasma with UV light.…”

Section: Irreversible Photo(de)activation (Uncaging)mentioning

confidence: 99%

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Light‐Controlled Tools

et al. 2012

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Spatial and temporal control over chemical and biological processes plays a key role in life, where the whole is often much more than the sum of its parts. Quite trivially, the molecules of a cell do not form a living system if they are only arranged in a random fashion. If we want to understand these relationships and especially the problems arising from malfunction, tools are necessary that allow us to design sophisticated experiments that address these questions. Highly valuable in this respect are external triggers that enable us to precisely determine where, when, and to what extent a process is started or stopped. Light is an ideal external trigger: It is highly selective and if applied correctly also harmless. It can be generated and manipulated with well-established techniques, and many ways exist to apply light to living systems--from cells to higher organisms. This Review will focus on developments over the last six years and includes discussions on the underlying technologies as well as their applications.

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Section: Irreversible Photo(de)activation (Uncaging)mentioning

confidence: 99%

“…Angewandte Chemie propionic acid, the isostere of cysteine. [131] By applying prothrombin as the extein in this system, they could induce coagulation in human blood plasma with UV light.…”

Section: Biochemical Photoswitchesmentioning

confidence: 99%

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et al. 2012

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“…Mootz et al stellten ein Intein mit NV-geschützter Diaminopropansäure her, die isoster zu Cystein ist. [131] Durch die Verwendung von Prothrombin als Extein konnten sie die Blutgerinnung in humanem Blutplasma mit Licht induzieren.…”

Section: Angewandte Aufsätzeunclassified

Lichtgesteuerte Werkzeuge

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Rohrbach

Gottschalk³

et al. 2012

Angewandte Chemie

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Die zeitliche und räumliche Kontrolle über chemische und biologische Prozesse spielt eine Schlüsselrolle im Leben, wo das Ganze oft viel mehr ist als die Summe seiner Einzelteile. Trivialerweise bilden die Moleküle einer Zelle noch lange kein lebendiges System aus, wenn sie lediglich willkürlich angeordnet werden. Wenn wir diese Zusammenhänge und die Probleme, die durch Fehlfunktionen entstehen, verstehen wollen, benötigen wir Werkzeuge, mit denen wir komplexe Experimente für diese Fragestellungen entwickeln können. Externe Triggersignale, mit denen wir präzise bestimmen können, wo, wann und in welchem Ausmaß ein Prozess gestartet oder gestoppt wird, sind dafür äußerst wertvoll. Licht ist solch ein ideales Triggersignal: Es ist hoch selektiv und bei richtiger Anwendung unschädlich. Es kann mit gut etablierten Techniken erzeugt und manipuliert werden, und viele Ansätze existieren, um Licht in lebenden Systemen anzuwenden – von Zellen bis zu höheren Organismen. Dieser Aufsatz wird sich mit den aktuellen Entwicklungen der letzten sechs Jahre befassen und die zugrundeliegenden Technologien sowie ihre Anwendungen diskutieren.

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“…[16] Other approaches explored split intein systems in which one of the fragments was chemically synthesized and contained a photochemically labile protection group positioned in a way that only the deprotected fragment gave rise to intein activity. [17] However, none of the mentioned systems has been used for the in vitro generation of cyclic peptides, partly because a two-fragment split intein strategy is not compatible with the SICLOPPS method for making cyclic peptide libraries.…”

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confidence: 99%

Generation of a Genetically Encoded, Photoactivatable Intein for the Controlled Production of Cyclic Peptides

et al. 2014

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Cyclic peptides are important natural products and hold great promise for the identification of new bioactive molecules. The split-intein-mediated SICLOPPS technology provides a generic access to fully genetically encoded head-to-tail cyclized peptides and large libraries thereof (SICLOPPS=split-intein circular ligation of peptides and proteins). However, owing to the spontaneous protein splicing reaction, product formation occurs inside cells, making peptide isolation inconvenient and precluding traditional in vitro assays for inhibitor discovery. The design of a genetically encoded, light-dependent intein using the photocaged tyrosine derivative ortho-nitrobenzyltyrosine incorporated at an internal, non-catalytic position is now reported. Stable intein precursors were purified from the E.coli expression host and subsequently subjected to light activation in vitro for both the regular protein splicing format and cyclic peptide production, including the natural product segetalin H as an example. The activity of the intein could also be triggered in living cells.

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Photocontrol of Protein Activity Mediated by the Cleavage Reaction of a Split Intein

Cited by 5 publications

References 27 publications

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Generation of a Genetically Encoded, Photoactivatable Intein for the Controlled Production of Cyclic Peptides

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