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DOI: 10.1039/C4CC05246C
Fig. 4: A – Enrichment of proteins labeled with β-keto ester probe. B – Enrichment of lysates spiked with alkynylated albumin. C – Comparative enrich-
ment of lysates alkynylated by probe 4 and enriched with linkers 5 and 6/streptavidin beads (SB) and azido-azo beads 2 (AAB).
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with the azo-moiety or other chemical groups in the protein-resin
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15 complex (ESI,† Fig. S3). The beads are not compatible with high
temperatures, which can lead to gelation of the beads.
In this work we have described the synthesis and biological
evaluation of a cleavable azide resin that can be used to isolate
alkyne-labeled proteins of virtually any size, making it useful for
20 general proteomics experiments e.g. ABPP. The enrichment
process is simplified requiring fewer precipitation and incubation
steps, and contamination of samples by naturally biotinylated
proteins and streptavidin subunits from the resin itself is avoided.
Furthermore, the cost is reduced since streptavidin beads are not
25 used. The synthesis requires no more work than that of a standard
cleavable biotin-linker and can be carried out on a large scale. For
these reasons we believe that azido-azo beads can be highly
useful for chemical biological applications and in the future
substitute the classical two step approach.
60
65
70
30
This work was financially supported by Aarhus University, the
Carlsberg Foundation, The Danish Council for Independent
Research, Medical and Natural Sciences (FSS and FNU).
Notes and references
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a Department of Chemistry, Aarhus University, Langelandsgade 140,
35 8000 Aarhus C, Denmark. E-mail: kaj@chem.au.dk
b Department of Forensic Medicine, Bioanalytical Unit, Aarhus Universi-
ty, Brendstrupgaardsvej 100, 8200 Aarhus N, Denmark. Tel: +45
87168332; E-mail: mj@forensic.au.dk
c Research Unit for Molecular Medicine, Institute of Clinical Medicine,
40 Aarhus University Hospital, Brendstrupgaardsvej 100, 8200 Aarhus N,
Denmark. E-mail: nig@ki.au.dk
† Electronic Supplementary Information (ESI) available: [details of any
supplementary information available should be included here]. See
DOI: 10.1039/b000000x/
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