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Single-site glycine-specific labeling of proteins

Author

Listed:
  • Landa Purushottam

    (Indian Institute of Science Education and Research (IISER) Bhopal)

  • Srinivasa Rao Adusumalli

    (Indian Institute of Science Education and Research (IISER) Bhopal)

  • Usha Singh

    (Indian Institute of Science Education and Research (IISER) Bhopal)

  • V. B. Unnikrishnan

    (Indian Institute of Science Education and Research (IISER) Bhopal)

  • Dattatraya Gautam Rawale

    (Indian Institute of Science Education and Research (IISER) Bhopal)

  • Mansi Gujrati

    (Indian Institute of Science Education and Research (IISER) Bhopal)

  • Ram Kumar Mishra

    (Indian Institute of Science Education and Research (IISER) Bhopal)

  • Vishal Rai

    (Indian Institute of Science Education and Research (IISER) Bhopal)

Abstract

Labeling of native proteins invites interest from diverse segments of science. However, there remains the significant unmet challenge in precise labeling at a single site of a protein. Here, we report the site-specific labeling of natural or easy-to-engineer N-terminus Gly in proteins with remarkable efficiency and selectivity. The method generates a latent nucleophile from N-terminus imine that reacts with an aldehyde to deliver an aminoalcohol under physiological conditions. It differentiates N-Gly as a unique target amongst other proteinogenic amino acids. The method allows single-site labeling of proteins in isolated form and extends to lysed cells. It administers an orthogonal aldehyde group primed for late-stage tagging with an affinity tag, 19F NMR probe, and a fluorophore. A user-friendly protocol delivers analytically pure tagged proteins. The mild reaction conditions do not alter the structure and function of the protein. The cellular uptake of fluorophore-tagged insulin and its ability to activate the insulin-receptor mediated signaling remains unperturbed.

Suggested Citation

  • Landa Purushottam & Srinivasa Rao Adusumalli & Usha Singh & V. B. Unnikrishnan & Dattatraya Gautam Rawale & Mansi Gujrati & Ram Kumar Mishra & Vishal Rai, 2019. "Single-site glycine-specific labeling of proteins," Nature Communications, Nature, vol. 10(1), pages 1-9, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-10503-7
    DOI: 10.1038/s41467-019-10503-7
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    Cited by:

    1. Neelesh C. Reddy & Rajib Molla & Pralhad Namdev Joshi & Sajeev T. K. & Ipsita Basu & Jyotsna Kawadkar & Neetu Kalra & Ram Kumar Mishra & Suman Chakrabarty & Sanjeev Shukla & Vishal Rai, 2022. "Traceless cysteine-linchpin enables precision engineering of lysine in native proteins," Nature Communications, Nature, vol. 13(1), pages 1-15, December.

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