title: Micropatterning proteins on polyhydroxyalkanoate substrates by using the substrate binding domain as a fusion partner.
abstract: A novel strategy for micropatterning proteins on the surface of polyhydroxyalkanoate (PHA) biopolymer by microcontact printing (microCP) 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.
authors: Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, Yuseong-gu, Daejeon, Republic of Korea.
authors: Jong Pil
authors: JP
authors: Park
authors: Kyung-Bok
authors: KB
authors: Lee
authors: Seok Jae
authors: SJ
authors: Lee
authors: Tae Jung
authors: TJ
authors: Park
authors: Min Gon
authors: MG
authors: Kim
authors: Bong Hyun
authors: BH
authors: Chung
authors: Zee-Won
authors: ZW
authors: Lee
authors: Insung S
authors: IS
authors: Choi
authors: Sang Yup
authors: SY
authors: Lee
mesh_terms: D000327:Adsorption; D001665:Binding Sites; D002265:Carboxylic Ester Hydrolases / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D020099:Coated Materials, Biocompatible / Q000032:analysis / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000032:analysis / Q000737:chemistry / Q000648:ultrastructure; D008422:Materials Testing; D011108:Polymers / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D044223:Pseudomonas stutzeri / Q000201:enzymology* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D013499:Surface Properties
chemical_list: D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D011108:Polymers; D011993:Recombinant Fusion Proteins; C111948:poly(hydroxyethanoate); D002265:Carboxylic Ester Hydrolases; C077226:poly(3-hydroxyalkanoic acid) depolymerase