Protein microarray using alpha-amino acids as metal tags on chips.
Procedures for synthesizing alpha-amino acids on a chip for coordination with transitional metal ions and a His-Tagged protein have been successfully developed as a stable protein microarray. Using the recombinant His-Tagged 3CL-protease (3CLpro) as a model for attachment to chips containing D-/L-Glu, Asp, Orn, Ser via different transitional metal ions, it was found that the Orn chip was the best of affinity binding and stability by which Zn2+ was the best metal ion for affinity while Co2+ was the best metal ion for stability. Thus, this protein microarray can be alternatively used as a high throughput screening method for rapid detection against SARS CoV 3CLpro and/or efficient purification of other Tagged proteins.
Sinchaikul
Supachok
S
Institute of Biological Chemistry and the Genomics Research Center, Academia Sinica, Nankang, Taipei 115, Taiwan.
Pan
Fu-Ming
FM
Cheng
Ching-Wen
CW
Wong
Chi-Huey
CH
Fang
Jim-Min
JM
Tseng
Min-Jen
MJ
Chen
Shui-Tein
ST
D000596:Amino Acids / Q000737:chemistry*; D019216:Metals, Heavy / Q000737:chemistry*; D008968:Molecular Conformation; D040081:Protein Array Analysis / Q000379:methods*; D011994:Recombinant Proteins / Q000032:analysis; D013329:Structure-Activity Relationship; D013997:Time Factors
D000596:Amino Acids; D019216:Metals, Heavy; D011994:Recombinant Proteins