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. 2010 Mar 8;49(11):2023-7.
doi: 10.1002/anie.200906756.

Bisaryl hydrazones as exchangeable biocompatible linkers

Anouk Dirksen  1 , Subramanian Yegneswaran, Philip E Dawson
Affiliations

Bisaryl hydrazones as exchangeable biocompatible linkers

Anouk Dirksen et al. Angew Chem Int Ed Engl. .
No abstract available

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Figures

Figure 1
Figure 1
(a) Biotinylation agent CLB354.[10] (b) CLB354 (5 μM) in 0.1 M Na phosphate (pH 6.0), in the absence of amines (stable), in presence of 100 mM aniline (reequilibration), and in the presence of 100 mM NH2OH and 100 mM aniline (capture).
None
Scheme 1
Three distinct strategies of cleaving a hydrazone linker: A by capping the protein, B by label exchange for further analysis or purification and C by capping the beads, leaving a synthetic handle on the protein for further modification.
None
Scheme 2
Retrieval of CLB354-labeled human serum albumin (HSA) from a well-defined protein mixture. Cleavage is achieved by capping the aldehyde on HSA with NH2OH (bottom, left) or the hydrazinopyridine group on biotin with PEGylated benzaldehyde (bottom, right). Fluorescently labeled HSA was obtained by cleavage with aminooxyacetyl-Alexa Fluor 488, whereas a FLAG-tag could be introduced by cleavage with aminooxyacetyl-FLAG (bottom, middle). Aniline catalyzes transimination and improves cleavage efficiencies. SDS-PAGE analysis (4-12% Bis-Tris gel, Simply Blue™ stain) shows modified HSA. The hydrazone linker is stable in pH 6.0 buffer (top, right). An SDS boil was performed as a reference (top, right). No detectable levels of the other proteins in the protein mixture are observed after elution. See SI for details.
See this image and copyright information in PMC

References

    1. Green NM. Methods Enzymol. 1990;184:51–67. - PubMed
    1. Hirsch JD, Eslamizar L, Filanoski BJ, Malekzadeh N, Haugland RP, Beechem JM, Haugland RP. Anal Biochem. 2002;308:343–357. - PubMed
    2. Zeheb R, Orr GA. Methods Enzymol. 1986;122:87–94. - PubMed
    3. Howarth M, Chinnapen DJF, Gerrow K, Dorrestein PC, Grandy MR, Kelleher NL, El-Husseini A, Ting AY. Nat Methods. 2006;3:267–273. - PMC - PubMed
    4. Wu SC, Wong SL. J Biol Chem. 2005;280:23225–23231. - PubMed
    5. Malmstadt N, Hyre DE, Ding Z, Hoffman AS, Stayton PS. Bioconjugate Chem. 2003;14:575–580. - PubMed
    6. Morag E, Bayer EA, Wilchek M. Biochem J. 1996;316:193–199. - PMC - PubMed
    1. Pfander S, Fiammengo R, Kirin SI, Metzler-Nolte N, Jäschke A. Nucleic Acids Res. 2007;35:e25. - PMC - PubMed
    2. Speers AE, Cravatt BF. J Am Chem Soc. 2005;127:10018–10019. - PMC - PubMed
    1. Verhelst SHL, Fonović M, Bogyo M. Angew Chem. 2007;119:1306–1308. Angew. Chem. Int. Ed. 2007, 46, 1284–1286. - PubMed
    2. Gartner CA, Elias JE, Bakalarski CE, Gygi SP. J Prot Res. 2007;6:1482–1491. - PubMed
    3. Fauq AH, Kache R, Kahn MA, Vega IE. Bioconjugate Chem. 2006;17:248–254. - PubMed
    4. van der Veken P, Dirksen EHC, Ruijter R, Elgersma RC, Heck AJR, Rijkers DTS, Slijper M, Liskamp RMJ. ChemBioChem. 2005;6:2271–2280. - PubMed
    5. Shimkus M, Levy J, Herman T. Proc Nat Acad Sci USA. 1985;82:2593–2597. - PMC - PubMed
    1. King TP, Zhao SW, Lam T. Biochemistry. 1986;25:5774–5779. - PubMed

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