Chemoselective immobilization of proteins by microcontact printing and bio-orthogonal click reactions

Zachary P Tolstyka, Wade Richardson, Erhan Bat, Caitlin J. Stevens, Dayanara P. Parra, Jonathan K. Dozier, Mark D Distefano, Bruce Dunn, Heather D. Maynard

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Herein, a combination of microcontact printing of functionalized alkanethiols and site-specific modification of proteins is utilized to chemoselectively immobilize proteins onto gold surfaces, either by oxime- or copper-catalyzed alkyne-azide click chemistry. Two molecules capable of click reactions were synthesized, an aminooxy-functionalized alkanethiol and an azide-functionalized alkanethiol, and self-assembled monolayer (SAM) formation on gold was confirmed by IR spectroscopy. The alkanethiols were then individually patterned onto gold surfaces by microcontact printing. Site-specifically modified proteins - horse heart myoglobin (HHMb) containing an N-terminal α-oxoamide and a red fluorescent protein (mCherry-CVIA) with a C-terminal alkyne - were immobilized by incubation onto respective stamped functionalized alkanethiol patterns. Pattern formation was confirmed by fluorescence microscopy. Don't move! Chemoselective immobilization of site-specifically modified proteins on surfaces through bio-orthogonal click reactions is demonstrated. The presented approach involves microcontact printing of aminooxy- or azide-functionalized alkanethiols and subsequent immobilization of proteins.

Original languageEnglish (US)
Pages (from-to)2464-2471
Number of pages8
JournalChemBioChem
Volume14
Issue number18
DOIs
StatePublished - Dec 1 2013

Fingerprint

Printing
Immobilization
Azides
Gold
Alkynes
Proteins
Click Chemistry
Oximes
Myoglobin
Fluorescence Microscopy
Horses
Fluorescence microscopy
Copper
Self assembled monolayers
Spectrum Analysis
Membrane Proteins
Infrared spectroscopy
Molecules

Keywords

  • chemoselectivity
  • click chemistry
  • microcontact printing
  • protein immobilization
  • self-assembly

Cite this

Tolstyka, Z. P., Richardson, W., Bat, E., Stevens, C. J., Parra, D. P., Dozier, J. K., ... Maynard, H. D. (2013). Chemoselective immobilization of proteins by microcontact printing and bio-orthogonal click reactions. ChemBioChem, 14(18), 2464-2471. https://doi.org/10.1002/cbic.201300478

Chemoselective immobilization of proteins by microcontact printing and bio-orthogonal click reactions. / Tolstyka, Zachary P; Richardson, Wade; Bat, Erhan; Stevens, Caitlin J.; Parra, Dayanara P.; Dozier, Jonathan K.; Distefano, Mark D; Dunn, Bruce; Maynard, Heather D.

In: ChemBioChem, Vol. 14, No. 18, 01.12.2013, p. 2464-2471.

Research output: Contribution to journalArticle

Tolstyka, ZP, Richardson, W, Bat, E, Stevens, CJ, Parra, DP, Dozier, JK, Distefano, MD, Dunn, B & Maynard, HD 2013, 'Chemoselective immobilization of proteins by microcontact printing and bio-orthogonal click reactions', ChemBioChem, vol. 14, no. 18, pp. 2464-2471. https://doi.org/10.1002/cbic.201300478
Tolstyka ZP, Richardson W, Bat E, Stevens CJ, Parra DP, Dozier JK et al. Chemoselective immobilization of proteins by microcontact printing and bio-orthogonal click reactions. ChemBioChem. 2013 Dec 1;14(18):2464-2471. https://doi.org/10.1002/cbic.201300478
Tolstyka, Zachary P ; Richardson, Wade ; Bat, Erhan ; Stevens, Caitlin J. ; Parra, Dayanara P. ; Dozier, Jonathan K. ; Distefano, Mark D ; Dunn, Bruce ; Maynard, Heather D. / Chemoselective immobilization of proteins by microcontact printing and bio-orthogonal click reactions. In: ChemBioChem. 2013 ; Vol. 14, No. 18. pp. 2464-2471.
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