Micropatterning proteins on polyhydroxyalkanoate substrates by using the substrate binding domain as a fusion partner.
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.
Park
Jong Pil
JP
Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, Yuseong-gu, Daejeon, Republic of Korea.
Lee
Kyung-Bok
KB
Lee
Seok Jae
SJ
Park
Tae Jung
TJ
Kim
Min Gon
MG
Chung
Bong Hyun
BH
Lee
Zee-Won
ZW
Choi
Insung S
IS
Lee
Sang Yup
SY
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
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