title: Traceless cross-linker for photocleavable bioconjugation.
abstract: Photoresponsive bioconjugation empowers the development of novel methods for drug discovery, disease diagnosis, and high-throughput screening, among others. In this paper, we report on the characteristics of a traceless photocleavable cross-linker, di-6-(3-succinimidyl carbonyloxymethyl-4-nitro-phenoxy)-hexanoic acid disulfide diethanol ester (SCNE). The traceless feature and the biocompatibility of this photocleavable cross-linking reagent were corroborated. Consequently, we demonstrated its application in reversible phage particle immobilization that could provide a platform for direct single-phage screening. We also applied it in protein-photoprinting, where SCNE acts as a "photo-eraser" to remove the cross-linked protein molecules at a desired region in a simple, clean, and light-controllable fashion. We further demonstrated the two-tier atomic force microscopic (AFM) method that uses SCNE to carry out two subsequent AFM tasks in situ. The approach allows guided protein delivery and subsequent high-resolution imaging at the same local area, thus opening up the possibility of monitoring protein functions in live cells. The results imply that SCNE is a versatile cross-linker that can be used for a wide range of applications where photocleavage ensures clean and remote-controllable release of biological molecules from a substrate.
authors.affiliation: Department of Biological, Chemical and Physical Sciences, Illinois Institute of Technology, Chicago, IL 60616, USA. wangr@iit.edu
authors.forename: Rong
authors.initials: R
authors.lastname: Wang
authors.forename: Funing
authors.initials: F
authors.lastname: Yan
authors.forename: Dengli
authors.initials: D
authors.lastname: Qiu
authors.forename: Jae-Sun
authors.initials: JS
authors.lastname: Jeong
authors.forename: Qiaoling
authors.initials: Q
authors.lastname: Jin
authors.forename: Tae-Young
authors.initials: TY
authors.lastname: Kim
authors.forename: Liaohai
authors.initials: L
authors.lastname: Chen
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D001435:Bacteriophages / Q000737:chemistry; D001672:Biocompatible Materials / Q000737:chemistry*; D002208:Caproates / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D006801:Humans; D018625:Microscopy, Atomic Force; D009574:Nitro Compounds / Q000737:chemistry*; D010782:Photolysis; D011327:Printing
chemical_list: D001426:Bacterial Proteins; D001672:Biocompatible Materials; D002208:Caproates; D003432:Cross-Linking Reagents; D009574:Nitro Compounds; C574333:di-6-(3-succinimidyl carbonyloxymethyl-4-nitro-phenoxy)-hexanoic acid disulfide diethanol ester; C037652:hexanoic acid