Protein microarrays to detect protein-protein interactions using red and green fluorescent proteins.
Proteomics, the study of protein function on a global scale, will play an important role in furthering our understanding of gene functions, complex biological pathways, and discovery of novel drug targets. A number of techniques have been developed for proteomic studies to identify and analyze proteins, compare protein expression levels, and study protein-protein interactions. Recent developments have applied a DNA array-type approach to immobilize proteins on a surface for high-throughput analysis. Here we report the development and construction of protein chips using derivatized glass and nitrocellulose-coated slides and the employment of recombinant proteins fused with green and red fluorescent proteins for detection. Fluorescent signals were found to be proportional to the amount of arrayed proteins and could be readily detected with a conventional fluorescence slide scanner. This technique allows the investigation of protein-protein interactions without the need for additional labeling steps of probe proteins.
Kukar
Thomas
T
Department of Medicinal Chemistry, College of Pharmacy, University of Florida, Gainesville, Florida 32610, USA.
Eckenrode
Sarah
S
Gu
Yunrong
Y
Lian
Wei
W
Megginson
Mike
M
She
Jin-Xiong
JX
Wu
Donghai
D
D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D004926:Escherichia coli; D005453:Fluorescence; D005898:Glass; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis*; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000032:analysis; D000097573:Red Fluorescent Protein
D000906:Antibodies; D000941:Antigens; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein