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An Intrinsically Fluorescent Recognition Ligand Scaffold Based on Chaperonin Protein and Semiconductor Quantum‐Dot Conjugates
Authors:Hongzhi Xie  Yi‐Fen Li  Hiromi K Kagawa  Jonathan D Trent  Kumara Mudalige  Mircea Cotlet  Basil I Swanson
Affiliation:1. Chemistry Division, Los Alamos National Laboratory Mail Stop J567, Los Alamos, NM 87545 (USA);2. Bioengineering Branch, NASA Ames Research Center Moffett Field, CA 94035 (USA);3. Center for Functional Nanomaterials, Brookhaven National Laboratory Mail Stop 735, Upton, NY 11973 (USA)
Abstract:Genetic engineering of a novel protein–nanoparticle hybrid system with great potential for biosensing applications and for patterning of various types of nanoparticles is described. The hybrid system is based on a genetically modified chaperonin protein from the hyperthermophilic archaeon Sulfolobus shibatae. This chaperonin is an 18‐subunit double ring, which self‐assembles in the presence of Mg ions and ATP. Described here is a mutant chaperonin (His‐β‐loopless, HBLL) with increased access to the central cavity and His‐tags on each subunit extending into the central cavity. This mutant binds water‐soluble semiconductor quantum dots, creating a protein‐encapsulated fluorescent nanoparticle. The new bioconjugate has high affinity, in the order of strong antibody–antigen interactions, a one‐to‐one protein–nanoparticle stoichiometry, and high stability. By adding selective binding sites to the solvent‐exposed regions of the chaperonin, this protein–nanoparticle bioconjugate becomes a sensor for specific targets.
Keywords:nanoparticles  proteins  quantum dots  semiconductors
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