IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v15y2024i1d10.1038_s41467-024-55212-y.html
   My bibliography  Save this article

Protein purification with light via a genetically encoded azobenzene side chain

Author

Listed:
  • Peter Mayrhofer

    (Technical University of Munich)

  • Markus R. Anneser

    (Technical University of Munich)

  • Kristina Schira

    (Technical University of Munich)

  • Carina A. Sommer

    (Technical University of Munich)

  • Ina Theobald

    (Technical University of Munich)

  • Martin Schlapschy

    (Technical University of Munich)

  • Stefan Achatz

    (Technical University of Munich)

  • Arne Skerra

    (Technical University of Munich)

Abstract

Affinity chromatography is the method of choice for the rapid purification of proteins from cell extracts or culture supernatants. Here, we present the light-responsive Azo-tag, a short peptide comprising p-(phenylazo)-L-phenylalanine (Pap), whose side chain can be switched from its trans-ground state to the metastable cis-configuration by irradiation with mild UV light. Since only trans-Pap shows strong affinity to α-cyclodextrin (α-CD), a protein exhibiting the Azo-tag selectively binds to an α-CD chromatography matrix under daylight or in the dark but elutes quickly under physiological buffer flow when illuminating the column at 355 nm. We demonstrate the light-controlled single-step purification – termed Excitography – of diverse proteins, including enzymes and antibody fragments, without necessitating competing agents or harsh buffer conditions as normally applied. While affinity chromatography has so far been governed by chemical interactions, introducing control by electromagnetic radiation as a physical principle adds another dimension to this widely applied separation technique.

Suggested Citation

  • Peter Mayrhofer & Markus R. Anneser & Kristina Schira & Carina A. Sommer & Ina Theobald & Martin Schlapschy & Stefan Achatz & Arne Skerra, 2024. "Protein purification with light via a genetically encoded azobenzene side chain," Nature Communications, Nature, vol. 15(1), pages 1-18, December.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-55212-y
    DOI: 10.1038/s41467-024-55212-y
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-024-55212-y
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-024-55212-y?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    References listed on IDEAS

    as
    1. Hiroyasu Yamaguchi & Yuichiro Kobayashi & Ryosuke Kobayashi & Yoshinori Takashima & Akihito Hashidzume & Akira Harada, 2012. "Photoswitchable gel assembly based on molecular recognition," Nature Communications, Nature, vol. 3(1), pages 1-5, January.
    2. Sherry K. Blight & Ross C. Larue & Anirban Mahapatra & David G. Longstaff & Edward Chang & Gang Zhao & Patrick T. Kang & Kari B. Green-Church & Michael K. Chan & Joseph A. Krzycki, 2004. "Direct charging of tRNACUA with pyrrolysine in vitro and in vivo," Nature, Nature, vol. 431(7006), pages 333-335, September.
    Full references (including those not matched with items on IDEAS)

    Most related items

    These are the items that most often cite the same works as this one and are cited by the same works as this one.
    1. Changyong Cai & Shuanggen Wu & Yunfei Zhang & Fenfang Li & Zhijian Tan & Shengyi Dong, 2024. "Bulk transparent supramolecular glass enabled by host–guest molecular recognition," Nature Communications, Nature, vol. 15(1), pages 1-9, December.
    2. Pang Zhu & Qingchuan Song & Sagar Bhagwat & Fadoua Mayoussi & Andreas Goralczyk & Niloofar Nekoonam & Mario Sanjaya & Peilong Hou & Silvio Tisato & Frederik Kotz-Helmer & Dorothea Helmer & Bastian E. , 2024. "Generation of precision microstructures based on reconfigurable photoresponsive hydrogels for high-resolution polymer replication and microoptics," Nature Communications, Nature, vol. 15(1), pages 1-10, December.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-55212-y. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    If CitEc recognized a bibliographic reference but did not link an item in RePEc to it, you can help with this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.