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Micropatterning proteins on polyhydroxyalkanoate substrates by using the substrate binding domain as a fusion partner
Author(s) -
Park Jong Pil,
Lee KyungBok,
Lee Seok Jae,
Park Tae Jung,
Kim Min Gon,
Chung Bong Hyun,
Lee ZeeWon,
Choi Insung S.,
Lee Sang Yup
Publication year - 2005
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20581
Subject(s) - micropatterning , fusion protein , substrate (aquarium) , microcontact printing , green fluorescent protein , surface plasmon resonance , polyhydroxyalkanoates , biopolymer , chemistry , fluorescence , biophysics , biochemistry , materials science , nanotechnology , biology , polymer , recombinant dna , bacteria , organic chemistry , nanoparticle , ecology , genetics , physics , quantum mechanics , gene
A novel strategy for micropatterning proteins on the surface of polyhydroxyalkanoate (PHA) biopolymer by microcontact printing (µCP) is described. The substrate binding domain (SBD) of the Pseudomonas stutzeri PHA depolymerase was used as a fusion partner for specifically immobilizing proteins on PHA substrate. Enhanced green fluorescent protein (EGFP) and red fluorescent protein (RFP) fused to the SBD could be specifically immobilized on the micropatterns of poly(3‐hydroxybutyrate) and poly(3‐hydroxybutyrate‐co‐3‐hydroxyhexanoate). Laser scanning confocal microscopic studies suggested that two fusion proteins were micropatterned in their functionally active forms. Also, antibody binding assay by surface plasmon resonance suggested that protein–protein interaction studies could be carried out using this system. © 2005 Wiley Periodicals, Inc.