Imidazole carbamate probes for affinity guided azide-transfer to metal-binding proteins

Michael Rosholm Mortensen; Nanna Louise Nielsen; Johan Palmfeldt; Kurt Vesterager Gothelf

doi:10.1039/C8OB03017K

Imidazole carbamate probes for affinity guided azide-transfer to metal-binding proteins

Michael Rosholm Mortensen, Nanna Louise Nielsen, Johan Palmfeldt, Kurt Vesterager Gothelf^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

13 Citations (Scopus)

Abstract

Probes for affinity guided conjugation have shown great promise for the preparation of high-quality protein conjugates. However, such probes are often limited in their protein scope. Here, we demonstrate the synthesis and use of imidazole carbamate probes for affinity guided conjugation of an azide handle to a wide variety of metal binding proteins such as antibodies, enzymes, nanobodies and His₆-tagged proteins. The azide handles were used to link dyes to the proteins for gel analysis and binding studies. In a comparative study we find that the probe containing three NTA groups show higher selectivity than the probe containing two. In a mixture of proteins, the probe shows preference for labelling His₆-tagged proteins.

Original language	English
Journal	Organic and Biomolecular Chemistry
Volume	17
Issue	6
Pages (from-to)	1379-1383
Number of pages	5
ISSN	1477-0520
DOIs	https://doi.org/10.1039/C8OB03017K
Publication status	Published - 1 Jan 2019

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title = "Imidazole carbamate probes for affinity guided azide-transfer to metal-binding proteins",

abstract = "Probes for affinity guided conjugation have shown great promise for the preparation of high-quality protein conjugates. However, such probes are often limited in their protein scope. Here, we demonstrate the synthesis and use of imidazole carbamate probes for affinity guided conjugation of an azide handle to a wide variety of metal binding proteins such as antibodies, enzymes, nanobodies and His6-tagged proteins. The azide handles were used to link dyes to the proteins for gel analysis and binding studies. In a comparative study we find that the probe containing three NTA groups show higher selectivity than the probe containing two. In a mixture of proteins, the probe shows preference for labelling His6-tagged proteins.",

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Imidazole carbamate probes for affinity guided azide-transfer to metal-binding proteins. / Mortensen, Michael Rosholm; Nielsen, Nanna Louise; Palmfeldt, Johan et al.
In: Organic and Biomolecular Chemistry, Vol. 17, No. 6, 01.01.2019, p. 1379-1383.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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T1 - Imidazole carbamate probes for affinity guided azide-transfer to metal-binding proteins

AU - Mortensen, Michael Rosholm

AU - Nielsen, Nanna Louise

AU - Palmfeldt, Johan

AU - Gothelf, Kurt Vesterager

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Probes for affinity guided conjugation have shown great promise for the preparation of high-quality protein conjugates. However, such probes are often limited in their protein scope. Here, we demonstrate the synthesis and use of imidazole carbamate probes for affinity guided conjugation of an azide handle to a wide variety of metal binding proteins such as antibodies, enzymes, nanobodies and His6-tagged proteins. The azide handles were used to link dyes to the proteins for gel analysis and binding studies. In a comparative study we find that the probe containing three NTA groups show higher selectivity than the probe containing two. In a mixture of proteins, the probe shows preference for labelling His6-tagged proteins.

AB - Probes for affinity guided conjugation have shown great promise for the preparation of high-quality protein conjugates. However, such probes are often limited in their protein scope. Here, we demonstrate the synthesis and use of imidazole carbamate probes for affinity guided conjugation of an azide handle to a wide variety of metal binding proteins such as antibodies, enzymes, nanobodies and His6-tagged proteins. The azide handles were used to link dyes to the proteins for gel analysis and binding studies. In a comparative study we find that the probe containing three NTA groups show higher selectivity than the probe containing two. In a mixture of proteins, the probe shows preference for labelling His6-tagged proteins.

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JO - Organic and Biomolecular Chemistry

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Imidazole carbamate probes for affinity guided azide-transfer to metal-binding proteins

Abstract

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