Site-specific immobilization of proteins in a microarray using intein-mediated protein splicing.
One of the critical issues in the generation of a protein microarray lies in the choice of immobilization strategies, which ensure proteins are adhered to the glass surface while properly retaining their native biological activities. Herein, we report a bacterium-based, intein-mediated strategy to generate N-terminal cysteine-containing proteins which are then chemoselectively immobilized to a thioester-functionalized glass slide to generate the corresponding protein microarray. We also showed preliminary data of the strategy in a yeast host system.
Girish
Aparna
A
Department of Biological Sciences, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
Sun
Hongyan
H
Yeo
Dawn S Y
DS
Chen
Grace Y J
GY
Chua
Teck-Khiang
TK
Yao
Shao Q
SQ
D004591:Electrophoresis, Polyacrylamide Gel; D047668:Inteins; D040081:Protein Array Analysis; D011506:Proteins / Q000737:chemistry*
D011506:Proteins