Intein-mediated biotinylation of proteins and its application in a protein microarray.
We report here the first example using an intein-mediated expression system to generate biotinylated proteins suitable for immobilization onto avidin-functionalized glass slides. With this novel array, proteins are site-specifically immobilized on the glass surface and are able to retain their native activity. The advantage of the avidin/biotin linkage over his-tag/Ni-NTA strategies for protein immobilization is highlighted by its ability to withstand a variety of chemical conditions, which makes this new protein array compatible with most biological assays.
Lesaicherre
Marie-Laure
ML
Department of Chemistry and Department of Biological Sciences, National University of Singapore, 3 Science Drive 3, Singapore 117543, Republic of Singapore.
Lue
Rina Y P
RY
Chen
Grace Y J
GY
Zhu
Qing
Q
Yao
Shao Q
SQ
D000906:Antibodies / Q000737:chemistry; D019921:Biotinylation; D002352:Carrier Proteins / Q000737:chemistry; D005982:Glutathione Transferase / Q000737:chemistry; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry; D058619:Maltose-Binding Proteins; D009532:Nickel / Q000737:chemistry; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry
D000906:Antibodies; D002352:Carrier Proteins; D008164:Luminescent Proteins; D058619:Maltose-Binding Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D009532:Nickel; D005982:Glutathione Transferase