A fusion protein expression analysis using surface plasmon resonance imaging.
A surface plasmon resonance (SPR) imaging system was constructed and used to detect the affinity-tagged recombinant proteins expressed in Escherichia coli. With regards to model proteins, the hexahistidine-ubiquitin-tagged human growth hormone (His(6)-Ub-hGH), glutathione S-transferase-tagged human interleukin-6 (GST-hIL6), and maltose-binding protein-tagged human interleukin-6 (MBP-hIL6) expressed in E. coli were analyzed. The cell lysates were spotted on gold thin films coated with 11-mercaptoundecanol (MUOH)/dextran derivatized with Ni(II)-iminodiacetic acid (IDA-Ni(II)), glutathione, or cyclodextrin. After a brief washing of the gold chip, SPR imaging measurements were carried out in order to detect the bound affinity-tagged fusion proteins. Using this new approach, rapid high-throughput expression analysis of the affinity-tagged proteins were obtained. The SPR imaging protein chip system used to measure the expression of affinity-tagged proteins in a high-throughput manner is expected to be an attractive alternative to traditional laborious and time-consuming methods, such as SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blots.
Jung
Jin-Mi
JM
Korea Research Institute of Bioscience and Biotechnology, BioNanotechnology Research Center, P.O. Box 115, Yuseong, Daejeon 305-600, Republic of Korea.
Shin
Yong-Beom
YB
Kim
Min-Gon
MG
Ro
Hyeon-Su
HS
Jung
Hee-Tae
HT
Chung
Bong Hyun
BH
D000345:Affinity Labels; D002352:Carrier Proteins / Q000032:analysis / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005233:Fatty Alcohols / Q000819:agonists; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics; D013006:Growth Hormone / Q000032:analysis / Q000235:genetics; D006639:Histidine / Q000235:genetics; D006801:Humans; D015850:Interleukin-6 / Q000032:analysis / Q000235:genetics; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics; D013438:Sulfhydryl Compounds / Q000819:agonists; D020349:Surface Plasmon Resonance / Q000379:methods*; D025801:Ubiquitin / Q000235:genetics
C521238:11-mercaptoundecanol; D000345:Affinity Labels; D002352:Carrier Proteins; D005233:Fatty Alcohols; D015850:Interleukin-6; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D013438:Sulfhydryl Compounds; D025801:Ubiquitin; D006639:Histidine; D013006:Growth Hormone; D005982:Glutathione Transferase