title: A methodology for preparing nanostructured biomolecular interfaces with high enzymatic activity.
abstract: The development of a novel method for functionalizing nanopatterned surfaces with catalytically active proteins is reported. This method involves using dip-pen nanolithography (DPN) and polymer pen lithography (PPL) to generate nanoscale patterns of coenzyme A, followed by a phosphopantetheinyl transferase-mediated coupling between coenzyme A and proteins fused to the ybbR-tag. By exploiting the ability to generate protein features over large areas afforded by DPN and PPL, it was now possible to measure protein activity directly on these surfaces. It was found that proteins immobilized on the nanoscale features not only display higher activity per area with decreasing feature size, but are also robust and can be used for repeated catalytic cycles. The immobilization method is applicable to a variety of proteins and gives rise to superior activity compared to proteins attached in random orientations on the surface.
authors.affiliation: Department of Chemistry and Institute of Nanotechnology, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208-3113, United States.
authors.forename: Lu Shin
authors.initials: LS
authors.lastname: Wong
authors.forename: Chinnan V
authors.initials: CV
authors.lastname: Karthikeyan
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Eichelsdoerfer
authors.forename: Jason
authors.initials: J
authors.lastname: Micklefield
authors.forename: Chad A
authors.initials: CA
authors.lastname: Mirkin
mesh_terms: D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis*; D003065:Coenzyme A / Q000737:chemistry* / Q000648:ultrastructure*; D016287:Computer Peripherals; D003460:Crystallization / Q000379:methods*; D004789:Enzyme Activation; D004800:Enzymes, Immobilized; D046911:Macromolecular Substances / Q000737:chemistry; D008422:Materials Testing; D044085:Microfluidics / Q000379:methods; D008968:Molecular Conformation; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure*; D010316:Particle Size; D013499:Surface Properties
chemical_list: D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D046911:Macromolecular Substances; D003065:Coenzyme A