title: Chemoselective immobilization of proteins by microcontact printing and bio-orthogonal click reactions.
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.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Los Angeles, 607 Charles E. Young Drive East, Los Angeles, CA 90095 (USA); California NanoSystems Institute, University of California, Los Angeles, 570 Westwood Plaza, Los Angeles, CA 90095 (USA).
authors.forename: Zachary P
authors.initials: ZP
authors.lastname: Tolstyka
authors.forename: Wade
authors.initials: W
authors.lastname: Richardson
authors.forename: Erhan
authors.initials: E
authors.lastname: Bat
authors.forename: Caitlin J
authors.initials: CJ
authors.lastname: Stevens
authors.forename: Dayanara P
authors.initials: DP
authors.lastname: Parra
authors.forename: Jonathan K
authors.initials: JK
authors.lastname: Dozier
authors.forename: Mark D
authors.initials: MD
authors.lastname: Distefano
authors.forename: Bruce
authors.initials: B
authors.lastname: Dunn
authors.forename: Heather D
authors.initials: HD
authors.lastname: Maynard
mesh_terms: D000473:Alkanes / Q000737:chemistry; D000480:Alkynes / Q000737:chemistry*; D000818:Animals; D001386:Azides / Q000737:chemistry*; D002384:Catalysis; D057930:Click Chemistry / Q000379:methods*; D003300:Copper / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006736:Horses; D055767:Immobilized Proteins / Q000737:chemistry*; D008164:Luminescent Proteins / Q000737:chemistry*; D009211:Myoglobin / Q000737:chemistry*; D010091:Oximes / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013499:Surface Properties; D000097573:Red Fluorescent Protein
chemical_list: D000473:Alkanes; D000480:Alkynes; D001386:Azides; D003300:Copper; D006046:Gold; D055767:Immobilized Proteins; D008164:Luminescent Proteins; D009211:Myoglobin; D010091:Oximes; D013438:Sulfhydryl Compounds; D000097573:Red Fluorescent Protein
keywords: chemoselectivity; click chemistry; microcontact printing; protein immobilization; self-assembly